Israeli scientists engineer plant seeds to produce milk protein without cows

Good news! Moo! Milk from seeds.

"... a new ... study has brought a step closer to the possibility that plant seeds could manufacture and store one of milk’s most important proteins – the same ones that give milk its nutrition, creamy texture, and cheese-making properties. ..."

From the abstract:

"... Casein is a high-quality protein source containing all amino acids, which are vital for human nutrition, and play a vital role in granting the texture and mouthfeel of dairy products.

Plant-seed production may be an efficient strategy for alternative protein production due to its ability to produce complex proteins, perform post-translational modifications, and be resource-efficient.

Targeting recombinant proteins to specific subcellular compartments plays a crucial role in ensuring proper folding, stability, and accumulation.

To determine the most suitable subcellular compartment for accumulation of bovine β-Casein fused to plant C-terminal Oleosin (CTO-Cas) different signal peptides were tested, directing the chimeric protein to endoplasmic-reticulum, vacuole or to the chloroplast.

These gene constructs were transformed into Arabidopsis using Agrobacterium-mediated transformation by the floral dip technique.

CTO-Cas was successfully detected in transformed seeds which were targeted to the vacuole, measured at 1.26% of the Total Soluble Protein (TSP). Subcellular compartmentation of the vacuole-targeted chimeric protein was determined by Transmitting Electron Microscopy (TEM) using gold-immuno-labeling.

CTO-Cas was detected in spherical bodies, which are not vacuoles.

Major morphological changes were observed in the transgenic seed cells in comparison to WT. Large oil bodies dominated WT seed cells, while seeds expressing CTO-Cas targeted to the vacuole had fewer large oil bodies. Instead, they exhibited numerous small oil bodies alongside dense sub-cellular aggregates only observed in the transgenic seeds.

These aggregates were positively gold-immunolabeled with anti-Casein antibodies as well as anti-oil body associated protein antibodies, suggesting the formation of recombinant CTO-Cas aggregates tightly associated with small oil bodies."

Israeli scientists engineer plants to produce milk protein without cows | The Jerusalem Post "The discovery would thus help overcome a major hurdle in producing real dairy proteins without cows, paving the way for more sustainable dairy ingredients and less climate change."

Microscope Reveals Surprising Milk Protein Clusters in Engineered Seeds (IMAGE)

Subcellular localization of C-term-oleosin fused to β-casein reveals unexpected cytoplasmic accumulation in vacuole-targeted arabidopsis seeds (open access)

Transmission electron microscopy (TEM) of engineered seed cells reveals that, instead of reaching its intended storage compartment, the milk protein formed previously unrecognized structures resembling natural casein micelles clustered near tiny oil bodies. Using gold nanoparticle labeling, researchers captured these novel, protein-rich clusters-visible as distinct dark circles-under high magnification to map their unexpected cellular location

Fig. 1 Schematic maps of the vector constructs used for Arabidopsis seeds genetic transformation, and the plasmid map (pCAMBIA 0309-BAR). (ER), endoplasmic reticulum; (Vac), vacuole; (Chl), chloroplast). Made with Bio Render.

The European Union is loosening its restrictions on certain gene-edited crops to take effect in two years. Really!

Good news, but too little, too late!

The excessive fear about genetic modifications in Europe is anachronistic and pathological!

"Last month, the European Parliament passed a law that will allow plants with genetic modifications that could occur naturally or through conventional breeding to bypass the EU’s onerous approval process for genetically modified organisms. The new regulatory process is expected to take effect in around two years."

"In a long-awaited step, the European Parliament yesterday passed a law that allows the expedited approval of many gene-edited crops without the rigorous review required for traditional transgenic varieties. ..."

Doomslayer: Progress Roundup - by Malcolm Cochran

New Law Lets EU More Quickly Greenlight Gene-Edited Crops

With new law, European Union can more quickly greenlight gene-edited crops

Implanted synthetically engineered bacteria to sense pathology and deliver drugs long-term to the site locally

Amazing stuff! This could be a breakthrough!

"Patient recovery from many debilitating conditions and diseases could be sped up significantly and be more effective if drugs and therapeutic molecules were delivered right to where they are needed in the body, over the entire regenerative process, and in doses finely tuned to therapeutic needs. An intriguing way to achieve this is the use of implantable, synthetically engineered, living cells that can sense injury or disease-associated conditions in their environment and flexibly respond by producing the right amount of a therapeutic molecule.

Bacteria, in particular, are promising in this regard as they can thrive in harsh physiological environments within the body, such as within infected or inflamed tissues, tissues undergoing mechanical movements, and tumors.

Some of these microbial therapies have even advanced into clinical trials to treat certain cancers, metabolic disorders, and the progression of kidney stones. However, thus far, such trials have failed, and microbes are feared to also pose significant safety risks because they cannot be contained at specific sites in the body. ...

By encapsulating a genetically engineered, therapeutic strain of E. coli bacteria within a biomaterial made from a hydrogel that was specifically designed to regulate bacterial growth and resist mechanical stresses, like those present at physically active sites in the body, the bacteria could be confined for over six months.

The E. coli bacteria were equipped with a synthetic gene circuit that allowed them to sense pathogenic Pseudomonas aeruginosa bacteria causing infections and then respond by releasing a therapeutic molecule that killed the nearby residing pathogens.

Implanted into the joints of mice next to a specialized orthopedic implant designed to help heal femoral injuries, the ILM autonomously and effectively treated infections with P. aeruginosa, a common cause of often debilitating orthopedic device infections. ..."

From the abstract of the Perspective:

"Engineered cells can sense disease and deliver drugs at a site of pathology. These living therapeutics provide localized, self-sustaining responses to environmental changes, such as inflammation and pathogenic signals, that conventional drugs cannot offer ... 

A promising chassis for living therapeutics is bacteria, which can be genetically programmed to release drugs in response to an external signal. However, bacteria require physical enclosure to prevent uncontrolled spread and toxicity. Biomaterials such as hydrogels and core-shell capsules have only demonstrated short-term containment of up to 2 weeks in culture. 

On page 729 of this issue, Harimoto et al. report a hydrogel scaffold with engineered stiffness and toughness that confines bacteria for up to 6 months in culture. When the system harbored bacteria producing pyocin, it cleared an infection in a mouse model of joint replacement. This could advance living therapeutics from short-lived proof-of-concept systems to durable, programmable medicines."

From the editor's summary and abstract:

"Editor’s summary

Engineered bacteria could serve as a source of long-term drug delivery, but they tend to escape confinement because of their small size and robust viability. Harimoto et al. created a polyvinyl alcohol (PVA) hydrogel matrix engineered for both high stiffness and high toughness that can contain bacteria without killing them off ... The hydrogel is used to trap engineered Escherichia coli that expresses a sense-and-respond genetic circuit designed to trigger the release of a protein antibiotic to clear Pseudomonas infection. This system was tested in vivo over a 6-month period, revealing positive treatment outcomes in a murine joint infection model. ...

Abstract

Microbes are increasingly used as living therapeutics, yet their uncontrolled dissemination in the body has remained a clinical roadblock.

Physical containment remains largely unattainable owing to eventual bacteria escape.

In this work, we present an implantable material that encapsulates and confines bacteria, wherein synthetically engineered microbes produce therapeutic payloads from within.

We developed a hydrogel scaffold with dual mechanical features: high stiffness to regulate bacterial proliferation and high toughness to resist material fracture under physiological stress.

This design achieved complete bacterial containment for 6 months and withstood multiple forms of mechanical loading that otherwise caused catastrophic material failure.

By genetically engineering embedded bacteria, we endowed the material with environmental sensing and on-demand therapeutic release capabilities and demonstrated autonomous treatment in a murine prosthetic joint infection model."

ScienceAdviser

Materializing safe, on-demand living therapeutics (original news release)

Scaffolds toughen bacteria-based therapy (Perspective, no public access)

Implantable living materials autonomously deliver therapeutics using contained engineered bacteria (no public access)

Implantable Living Materials Autonomously Deliver Therapeutics from Contained Engineered Bacteria (preprint, open access)

This illustration explains how the team designed Implantable Living Materials (ILMs) as a living therapeutic that uses an optimized hydrogel to safely contain synthetically engineered bacteria that are able to sense a pathogenic stimulus and respond to it by secreting a therapeutic protein within living organisms. The material itself is sufficiently “stiff” so that bacteria pushing against it from the inside can’t break it apart, and sufficiently “tough” to provide to protect the enclosed bacteria against external physical stresses. Combined with the synthetically engineered bacteria, the new approach becomes a safe and autonomous functioning drug delivery device.

Fig. 1 Design and mechanical characterization of Implantable Living Materials (ILMs).

Fig. 2 ILMs maintain bacterial containment during long-term culture and mechanical loading.

Scientists arrive at minimal, viable plant genomes through large chromosomal deletions

Amazing stuff!

"... Ancient events in plant evolution have left behind large, duplicated regions in their genomes.

... scientists found that deleting these large blocks of DNA can still lead to normal plants.

The findings demonstrate that large chromosomal deletions are a viable strategy in plant genetic engineering, which could now accelerate the development of streamlined, minimal plant genomes—a major goal in industries looking to create new plant-based biotechnologies. ...

The researchers used CRISPR-Cas9 to delete four large, duplicated blocks in Arabidopsis thaliana, a model plant commonly used in plant biology research. The deletions were then verified using whole-genome sequencing, which revealed minimal off-target effects. ..."

From the significance and abstract:

"Significance

Plant genomes are shaped by ancient polyploidy events, leaving behind extensive duplicated regions whose biological roles remain largely uncharacterized.

In this study, we demonstrate that targeted deletion of large, retained syntenic blocks in Arabidopsis thaliana results in viable plants, some with distinct phenotypes and widespread transcriptomic changes.

These findings challenge the assumption that such regions are essential and highlight the potential redundancy or modularity within plant genomes. Our approach of removing entire duplicated blocks offers a powerful strategy to functionally dissect conserved genomic regions, investigate gene linkage and dosage effects, and accelerate the development of streamlined, minimal plant genomes. This work establishes a scalable framework for genome engineering with broad implications for plant biology, synthetic genomics, and biotechnology.

Abstract

Plant genomes have undergone multiple rounds of whole-genome duplication (WGD) throughout their evolutionary history. As a result, many species, including Arabidopsis thaliana, retain duplicated genomic segments, or syntenic regions, which harbor large numbers of paralogous genes preserved from these ancient WGD events.

We deleted four large, duplicated blocks, ranging from ~115 kb to ~684 kb using Staphylococcus aureus Cas9 to explore the effects of knocking out these blocks in Arabidopsis.

Large deletions like these remain rare, especially in small and gene-dense plant genomes. Deletions were subsequently verified using whole-genome sequencing, which revealed minimal off-target effects.

The number of deleted genes ranged from 16 to 60, and transposable elements ranged from 4 to 112 among the four deleted blocks.

Two deletion lines showed distinct phenotypes resulting from the loss of many genes, while two others displayed no obvious defects, including for flowering time or hypocotyl elongation.

Moreover, RNA-sequencing revealed that expression compensation, where deletions of paralogous genes lead to the upregulation of intact paralogues, was not a general response to the deleted regions under the conditions tested.

Thus, it is possible to obtain viable plants when deleting large fragments that may be redundant or that contain nonessential genes.

These results demonstrate that large chromosomal deletions can be used as a tool for genome engineering approaches, such as genome minimization in plants and allele replacement using homology-directed repair and other precision editing methods. Targeted deletions of large chromosome fragments will be a valuable tool for research and biotechnology applications."

Scientists get back to basics with minimal plant genomes | EurekAlert!

Targeted deletions of large syntenic regions in Arabidopsis thaliana (open access)

Fig. 4 Generation of deletion lines (deletions of retained duplicate regions or nonlethal genomic segments) and their applications. 

Engineered bacteria produce ‘nylon’ for the first time

Amazing stuff! This could be a breakthrough!

"Polymers produced by bacteria might one day replace one of the most widely used fossil-fuel-based plastics: nylon. No natural enzymes produce this type of polymer, so researchers tweaked enzyme-coding genes from a variety of bacteria and inserted them into Escherichia coli. These genes then encoded several new-to-nature enzymes that could link up chains of molecules to create polymers, creating a bioplastic called poly(ester amide), or PEA. There are many hurdles to overcome before this laboratory experiment can be translated into a product: the PEA polymers have to be purified before they can be used, and the process is currently more expensive than the fossil-fuel route."

"Researchers have genetically engineered microbes to produce a strong, flexible plastic similar to nylon for the first time.

Bacteria have been used to generate polyesters such as polyhydroxyalkanoates (PHAs) in the past, but nylon-like plastics such as those used in clothing and shoe manufacturing have been difficult to create ...

Testing revealed that one type of PEA had physical, thermal and mechanical properties comparable to those of polyethylene, one of the most widely used commercial plastics. ..."

From the abstract:

"The development of biobased polymers to substitute their current petroleum-based counterparts is crucial for fostering a sustainable plastic industry.

Here we report the biosynthesis and characterization of a group of biopolymers, poly(ester amide)s (PEAs), in Escherichia coli. PEAs are biosynthesized by constructing a new-to-nature amino acid polymerization pathway, comprising amino acid activation by β-alanine CoA transferase and subsequent polymerization of amino acyl-CoA by polyhydroxyalkanoate synthase.

The engineered E. coli strains harboring this pathway are capable of biosynthesizing various PEAs, each incorporating different amino acid monomers in varying fractions.

Examination of the physical, thermal and mechanical properties reveals a dependence of molecular weight on the type of polyhydroxyalkanoate synthase, a decrease in melting temperature and crystallinity as the 3-aminopropionate monomer fraction increases and enhanced elongation at break compared to its polyester analog. The engineered bacterial system will prove beneficial for the biobased production of various PEAs using renewable resources."

Nature Briefing: Translational Research

Strong, flexible ‘nylon’ made by engineered bacteria for the first time "The bioplastic was malleable, but is more expensive to produce than are plastics made from fossil fuels."

Biosynthesis of poly(ester amide)s in engineered Escherichia coli (no public access)

Graphical abstract

Gene edited Insulin-producing beta cells avoid immune attack in treatment of type 1 diabetes over extended period

Good news!

"Gene-edited insulin-producing beta cells have survived for a month after being injected into a man with type 1 diabetes. These tweaked cells have a protein called CD47 on their surface, which tells the immune system not to destroy them. Only a small number of cells were injected into the man’s arm, but they have produced insulin and avoided his immune system’s crosshairs so far, suggesting that they could become an effective treatment for diabetes."

Nature Briefing: Translational Research

Gene-edited cells that evade rejection show promise in type 1 diabetes "Insulin-producing cells injected into a man with type 1 diabetes have survived for a month so far without the need for immune suppression"

Sana Biotechnology Announces Positive Clinical Results from Type 1 Diabetes Study of Islet Cell Transplantation Without Immunosuppression (original 

Remote-controlled gene therapy uses ultrasound to kill cancer

Good news! Amazing stuff! Cancer is history (soon)!

"... The resulting toolkit will allow CRISPR gene editing to be precisely targeted to the specific areas requiring treatment. The research team is already applying this discovery to refine their work in cancer immunotherapy. ...

But we are pushing it one step further to make it controllable. Instead of continuously editing the genome, we can now control it to be activated at a specific location and at a specific time using a non-invasive remote-controlled ultrasound wave. That’s the breakthrough.” ...

focused ultrasound as part of groundbreaking cancer immunotherapy research using engineered Chimeric Antigen Receptor (CAR) T-cells — immune cells that are extracted from patients and modified to be more effective against cancer. ... team harness ultrasound waves to directly control these CAR T-cells for precision targeting of tumor cells without harming healthy tissue. ...

“This is the first study that provides a very comprehensive, ultrasound-controllable CRISPR toolbox to knock out, activate, or silence a specific gene,” ...

“The telomere has many, many repeats, and we use CRISPR, guided by ultrasound, to cut this telomere so that it will trigger the chromosome to be cut off at two ends,”  ... “Because they have a repeatable sequence, they will all be cut, and therefore, the tumor cell can no longer repair itself. It’s all broken. The cell will then undergo apoptosis and die.” ..."

From the abstract:

"There remains a critical need for the precise control of CRISPR (clustered regularly interspaced short palindromic repeats)-based technologies. Here, we engineer a set of inducible CRISPR-based tools controllable by focused ultrasound (FUS), which can penetrate deep and induce localized hyperthermia for transgene activation. We demonstrate the capabilities of FUS-inducible CRISPR, CRISPR activation (CRISPRa), and CRISPR epigenetic editor (CRISPRee) in modulating the genome and epigenome. We show that FUS-CRISPR-mediated telomere disruption primes solid tumours for chimeric antigen receptor (CAR)-T cell therapy. We further deliver FUS-CRISPR in vivo using adeno-associated viruses (AAVs), followed by FUS-induced telomere disruption and the expression of a clinically validated antigen in a subpopulation of tumour cells, functioning as “training centers” to activate synthetic Notch (synNotch) CAR-T cells to produce CARs against a universal tumour antigen to exterminate neighboring tumour cells. The FUS-CRISPR(a/ee) toolbox hence allows the noninvasive and spatiotemporal control of genomic/epigenomic reprogramming for cancer treatment."

Remote-controlled gene therapy uses ultrasound to kill cancer

New CRISPR Toolkit to Allow Remote-Controlled Genome Editing (original news release) "USC Viterbi biomedical engineers harness focused ultrasound to revolutionize CRISPR’s capabilities to treat countless diseases."

Ultrasound Control of Genomic Regulatory Toolboxes for Cancer Immunotherapy (open access)

Fig. 4: FUS-CRISPR-mediated telomere disruption can inhibit tumor cell growth and its resistance to CAR-T cell killing.

Low-calorie sugar breakthrough could make allulose a household name

Good news!

"... One such substitute, allulose, is around 70% as sweet as sucrose, but contains just 10% of the calories and has even been shown to improve blood glucose levels and help in weight loss in people with type 2 diabetes. ...

Allulose – which is also known as D-psicose – is considered a rare sugar as it only exists naturally in minute amounts in a few plant foods, such as wheat, figs and raisins. When extracted, it has the texture and mouthfeel of sucrose, it has just 0.4 calories per gram, compared to four calories per gram in sucrose. And because it's a monosaccharide, or a single molecule of sugar, it undergoes a very different process in the body. Around 70% is absorbed by the small intestine and leaves the body via urine within 24 hours. The rest will exit the body ... through the large intestine, within about 48 hours. ...

The team ... edited the [E. coli] microorganism’s metabolic processes, so when the cells were fed glucose, they converted it to allulose. It immediately resulted in yields of 62% (and, importantly, a purity level in excess of 95%). ..."

From the abstract:

"Due to the rampant rise in obesity and diabetes, consumers are desperately seeking for ways to reduce their sugar intake, but to date there are no options that are both accessible and without sacrifice of palatability. One of the most promising new ingredients in the food system as a non-nutritive sugar substitute with near perfect palatability is D-psicose. D-psicose is currently produced using an in vitro enzymatic isomerization of D-fructose, resulting in low yield and purity, and therefore requiring substantial downstream processing to obtain a high purity product. This has made adoption of D-psicose into products limited and results in significantly higher per unit costs, reducing accessibility to those most in need. Here, we found that Escherichia coli natively possesses a thermodynamically favorable pathway to produce D-psicose from D-glucose through a series of phosphorylation-epimerization-dephosphorylation steps. To increase carbon flux towards D-psicose production, we introduced a series of genetic modifications to pathway enzymes, central carbon metabolism, and competing metabolic pathways. In an attempt to maximize both cellular viability and D-psicose production, we implemented methods for the dynamic regulation of key genes including clustered regularly interspaced short palindromic repeats inhibition (CRISPRi) and stationary-phase promoters. The engineered strains achieved complete consumption of D-glucose and production of D-psicose, at a titer of 15.3 g L-1, productivity of 2 g L-1 h-1, and yield of 62% under test tube conditions. These results demonstrate the viability of whole-cell catalysis as a sustainable alternative to in vitro enzymatic synthesis for the accessible production of D-psicose."

Low-calorie sugar breakthrough could make allulose a household name

Awakening the natural capability of psicose production in Escherichia coli (open access)

Fig. 1: Strategies for the biosynthesis of D-psicose.

Engineered Bacteria Tackle Pathogenic Biofilms of another bacteria in Mice to effectively treat lung infections

Amazing stuff! Good news! Ventilator associated pneumonia is particularly nasty: "The mortality in ventilator-associated pneumonia is high despite the availability of effective antibiotics." (Source)

"Mycoplasma pneumoniae are tiny bacteria typically known to cause lung infections. ... Genetically engineered Mycoplasma helped break down biofilms of another pathogenic microbe, Pseudomonas aeruginosa, in a mouse model of ventilator-associated pneumonia and on tube samples taken from human patients ... It is one of the first times that scientists have used live bacteria to treat a lung disease, and is the first therapeutic use of Mycoplasma. ...
Because of their complicated structure, biofilms are often resistant to antimicrobial treatments even when the individual bacteria in the film are not ...

chose Mycoplasma pneumoniae, a bacteria known for infecting the human respiratory system, for their experiments. Unlike many bacteria, M. pneumoniae cannot recombine, making it a safe candidate for bioengineering that won’t spread its modified genome to other bacteria. It also has a relatively small genome and lacks a cell wall, making it less likely to generate an immune reaction.

The researchers removed the pathogenic genes from M. pneumoniae to ensure the bacteria were safe to use. They then assembled sets of engineered genomes in E. coli, which scientists ... use to engineer bacterial treatments for the gut. These genomes contained a one-two punch to tackle the biofilm: a genetic cassette that codes for the production of biofilm-breaking enzymes and the gene for a toxin that can kill P. aeruginosa. They then transferred the genes in the form of isolated DNA to M. pneumoniae. ..."

From the abstract:

"Engineered live bacteria could provide a new modality for treating lung infections, a major cause of mortality worldwide. In the present study, we engineered a genome-reduced human lung bacterium, Mycoplasma pneumoniae, to treat ventilator-associated pneumonia, a disease with high hospital mortality when associated with Pseudomonas aeruginosa biofilms. After validating the biosafety of an attenuated M. pneumoniae chassis in mice, we introduced four transgenes into the chromosome by transposition to implement bactericidal and biofilm degradation activities. We show that this engineered strain has high efficacy against an acute P. aeruginosa lung infection in a mouse model. In addition, we demonstrated that the engineered strain could dissolve biofilms formed in endotracheal tubes of patients with ventilator-associated pneumonia and be combined with antibiotics targeting the peptidoglycan layer to increase efficacy against Gram-positive and Gram-negative bacteria. We expect our M. pneumoniae-engineered strain to be able to treat biofilm-associated infections in the respiratory tract."

Double Agents: Engineered Bacteria Tackle Pathogenic Biofilms in Mice | The Scientist Magazine® Mycoplasma pneumoniae with pathogenic genes replaced by biofilm-degrading ones enhance survival in a mouse model of ventilator-associated pneumonia.

Engineered live bacteria suppress Pseudomonas aeruginosa infection in mouse lung and dissolve endotracheal-tube biofilms

Fig. 5: In vivo treatment of mice with acute respiratory PAO1 infection.

CRISPR tools found in thousands of viruses could boost gene editing

Amazing stuff! CRISPR is everywhere it seems!

"A systematic sweep of viral genomes has revealed a trove of potential CRISPR-based genome-editing tools.

CRISPR–Cas systems are common in the microbial world of bacteria and archaea, where they often help cells to fend off viruses. But an analysis... finds CRISPR–Cas systems in 0.4% of publicly available genome sequences from viruses that can infect these microbes. Researchers think that the viruses use CRISPR–Cas to compete with one another — and potentially to manipulate gene activity in their host to their advantage. ..."

From the highlights and abstract:

"Highlights

• CRISPR pathways encoded in diverse bacteriophage are hypercompact anti-viral systems

• Phage-encoded CRISPR systems encompass all known CRISPR-Cas types

• The Casλ enzyme’s compact structure is capable of plant and human cell genome editing

• These findings reveal a new source of CRISPR-Cas enzymes with value as genome editors

Summary

CRISPR-Cas systems are host-encoded pathways that protect microbes from viral infection using an adaptive RNA-guided mechanism. Using genome-resolved metagenomics, we find that CRISPR systems are also encoded in diverse bacteriophages, where they occur as divergent and hypercompact anti-viral systems. Bacteriophage-encoded CRISPR systems belong to all six known CRISPR-Cas types, though some lack crucial components, suggesting alternate functional roles or host complementation. We describe multiple new Cas9-like proteins and 44 families related to type V CRISPR-Cas systems, including the Casλ RNA-guided nuclease family. Among the most divergent of the new enzymes identified, Casλ recognizes double-stranded DNA using a uniquely structured CRISPR RNA (crRNA). The Casλ-RNA-DNA structure determined by cryoelectron microscopy reveals a compact bilobed architecture capable of inducing genome editing in mammalian, Arabidopsis, and hexaploid wheat cells. These findings reveal a new source of CRISPR-Cas enzymes in phages and highlight their value as genome editors in plant and human cells."

CRISPR tools found in thousands of viruses could boost gene editing Phages probably picked up DNA-cutting systems from microbial hosts, and might use them to fight other viruses.

Diverse virus-encoded CRISPR-Cas systems include streamlined genome editors (open access)

Graphical abstract

Researchers Fuse Mouse Chromosomes in Scientific First

Amazing stuff!

"For the first time, researchers have fused two mouse chromosomes together in vitro, resulting in living mice with new karyotypes. ...
Chromosomal fusions are also common in cancer and have been linked to health issues, including infertility, aneuploidy, and childhood diseases. So, researchers have long sought the ability to precisely manipulate chromosomes in model organisms, especially mammalian ones, in the hopes of investigating fusions from both medical and evolutionary perspectives. ...
To fuse chromosomes in mice, the researchers used a technology they first developed in yeast: briefly, they injected modified haploid mouse embryonic stem cells friend (haESCs) with a CRISPR-Cas9 system that targets and eliminates telomeres and centromeres on two specific chromosomes. As a result, the targeted chromosomes zipped themselves together. ..."

"Designer chromosomes

One of the goals in synthetic biology is to generate complex multicellular life with designed DNA sequences. Being able to manipulate DNA at large scales, including at the chromosome level, is an important step toward this goal. So far, chromosome-level genetic engineering has been accomplished only in haploid yeast. By applying gene editing to haploid embryonic stem cells, Wang et al. achieved whole-chromosome ligations in mice, and successfully derived animals with 19 pairs of chromosomes, one pair fewer than is standard in this species."

From the abstract:

"Chromosome engineering has been attempted successfully in yeast but remains challenging in higher eukaryotes, including mammals. Here, we report programmed chromosome ligation in mice that resulted in the creation of new karyotypes in the lab. Using haploid embryonic stem cells and gene editing, we fused the two largest mouse chromosomes, chromosomes 1 and 2, and two medium-size chromosomes, chromosomes 4 and 5. Chromatin conformation and stem cell differentiation were minimally affected. However, karyotypes carrying fused chromosomes 1 and 2 resulted in arrested mitosis, polyploidization, and embryonic lethality, whereas a smaller fused chromosome composed of chromosomes 4 and 5 was able to be passed on to homozygous offspring. Our results suggest the feasibility of chromosome-level engineering in mammals."

Researchers Fuse Mouse Chromosomes in Scientific First | The Scientist Magazine®

A sustainable mouse karyotype created by programmed chromosome fusion (no public access)

Cancer-killing bacteria evade the immune system

Good news! Fascinating stuff! Interesting approach to treat cancer!

"... Over the past decade, researchers have explored the reduction of toxicities from live bacteria by genetically deleting the parts of the bacterium that can cause toxicity; but this can lead to unwanted mutations in the bacterium itself and may substantially decrease therapeutic efficacy.
A team of engineers ... has now determined an effective approach to enhance the delivery of living engineered bacteria into cells, while maintaining the bacterium’s integrity and minimizing toxicity. ... the researchers describe a way of coating engineered bacteria with an inducible capsular polysaccharide (iCAP) that responds in a smart manner when delivered into the body.
Capsular polysaccharide (CAP) is a layer of water molecules that coats the surface of natural bacteria and acts as a shield against foreign infections. By converting CAP into iCAP, the researchers could apply programmable external stimulus that enables the engineered bacteria to evade immune attack, survive for a considerable duration in the host environment and deliver a tolerable therapeutic dose. ..."

From the abstract:

"Living bacteria therapies have been proposed as an alternative approach to treating a broad array of cancers. In this study, we developed a genetically encoded microbial encapsulation system with tunable and dynamic expression of surface capsular polysaccharides that enhances systemic delivery. Based on a small RNA screen of capsular biosynthesis pathways, we constructed inducible synthetic gene circuits that regulate bacterial encapsulation in Escherichia coli Nissle 1917. These bacteria are capable of temporarily evading immune attack, whereas subsequent loss of encapsulation results in effective clearance in vivo. This dynamic delivery strategy enabled a ten-fold increase in maximum tolerated dose of bacteria and improved anti-tumor efficacy in murine models of cancer. Furthermore, in situ encapsulation increased the fraction of microbial translocation among mouse tumors, leading to efficacy in distal tumors. The programmable encapsulation system promises to enhance the therapeutic utility of living engineered bacteria for cancer."

Cancer-killing bacteria evade the immune system – Physics World

A programmable encapsulation system improves delivery of therapeutic bacteria in mice (open access)

"a, We engineered the biosynthetic pathway of bacterial CAP for tunable and dynamic surface modulation of the probiotic E. coli Nissle 1917 with synthetic gene circuits. This approach enables increased CAP levels upon induction to control immune evasion and clearance. b, The programmable CAP system enhances systemic delivery of bacteria by transiently expressing CAP. Non-CAP bacteria (thin gray cells) elicit toxicity by exposing the immunogenic bacterial surface, and permanently CAP-expressing bacteria (thick black cells) lead to overgrowth. The iCAP (blue cells) system enables transient encapsulation of bacteria, thus reducing initial inflammation while effectively clearing bacteria over time. c, The CAP system controls bacterial translocation among tumors. The iCAP system allows in situ activation of CAP in one tumor, which results in inducible bacteria translocation to distal, uncolonized tumors."

Lab-grown hair cells to treat baldness could be on the way

More snake oil or have researchers indeed finally come up with a solution that works? I have little doubt that we are coming closer to defeating baldness in men!

"... company says it can produce the components of hair follicles by genetically “reprogramming” ordinary cells, like blood or fat. Their hope is that the technology could eventually treat the underlying cause of hair loss. Given about half of men undergo male-pattern baldness, some starting in their 20s, there would be a substantial market. ..."

"Biologists at several startups are applying the latest advances in genetic engineering to the age-old problem of baldness, creating new hair-forming cells that could restore a person’s ability to grow hair. ...
In practice, though, the formula for producing specific cell types can prove elusive, and then there’s the problem of getting lab-grown cells back into the body. So far, there have been only a few demonstrations of reprogramming as a way to treat patients. Researchers in Japan tried transplanting retina cells into blind people. Then, last November, a US company, Vertex Pharmaceuticals, said it might have cured a man’s type 1 diabetes after an infusion of programmed beta cells, the kind that respond to insulin. ..."

Lab-grown hair cells to treat baldness could be on the way | MIT Technology Review These biotech companies are reprogramming cells to treat baldness, but it’s still early days.

A hairless mouse sprouts a tuft of human hair

following a transplant of follicle-forming stem cells

This DNA Factory Is Aiming to Reprogram the World

Recommendable! This is about Gingko biloba ahm  Ginkgo Bioworks! And Mofif Foodworks ... Artificial meet, restoring lost perfumes from extinct plants etc. However, this representative of Gingko Bioworks is awful!

UK to ease restrictions on gene editing in crops and livestock

Good news! The benefits of Brexit keep on giving!

"... Current regulations, inherited from the EU, class all gene-edited products as genetically-modified organisms (GMOs). This means that minor edits like gene deletions or single base-pair substitutions made to an organism’s genome are treated the same way legally as the introduction of DNA from other species. Many researchers argue that this fails to take into account scientific evidence and hinders the development of crops with desirable traits like disease-resistance and tolerance to drought. ..."

UK to ease restrictions on gene editing in crops and livestock | News | Chemistry World Redefinition of genetically-modified organisms could see Crispr-edited foods brought to market

Chinese Scientists Produce Genetically Modified Pigs for Human Transplant

Good news! Successful porcine xenotranplantation is coming another step or more closer! Lucky pig charm!

This research is not the latest. First published December 2019 as a preprint, then published September 2020 in Nature Biomedical Engineering. U.S. based companies and scientists were also involved.

"eGenesis [Cambridge, Massachusetts] is a gene editing and genome engineering company committed to the development of safe and effective human transplantable organs, tissues and cells to address the global organ crisis." (Source)

"... Xinhua reported that the team of Yang Luhan, founder of Qihan Biotech in Hanzhou, China, and cofounder and chief scientist of Cambridge gene-editing company eGenesis in Boston, had created a prototype xenograft with clinical potential, successfully solving two major xenograft safety challenges: removing porcine endogenous retroviruses from pigs and enhancing xenograft immunocompatibility. ...
In 2017, eGenesis announced that it had produced the world’s first genetically modified pigs that do not carry endogenous retroviruses, eliminating the risk of virus transmission from pigs to humans; and in 2018, eGenesis produced the first immunoplex-engineered pig, reducing the immune rejection of pig allogeneic organ transplants. ..."

"Xenotransplantation, specifically the use of porcine organs for human transplantation, has long been sought after as an alternative for patients suffering from organ failure. However, clinical application of this approach has been impeded by two main hurdles: 1) risk of transmission of porcine endogenous retroviruses (PERVs) and 2) molecular incompatibilities between donor pigs and humans which culminate in rejection of the graft. ...
In this study, we improved the scale of porcine germline editing from targeting a single repetitive locus with CRISPR to engineering 13 different genes using multiple genome engineering methods. ..."

Chinese Scientists Produce Genetically Modified Pigs for Human Transplant (behind paywall)

Extensive germline genome engineering in pigs (no public access)

Extensive Mammalian Germline Genome Engineering (open access, preprint)

https://arep.med.harvard.edu/pdf/Yue_Yang_2020.pdf

FDA approves genetically altering pigs, potentially for drugs or transplants

Good news! This is only the beginning! More to come!

"Genetically engineering pigs so they lack a certain sugar on the surface of their cells that triggers meat allergies or organ rejection won approval from the Food and Drug Administration Monday. The regulatory clearance — the first of an intentional genomic alteration in a product with both food and medical uses — means the animals could be safer sources of not just food but also treatments such as the blood-thinner heparin. ...
GalSafe pigs, named for their lack of detectable alpha-gal sugar, could potentially provide tissues and organs for patients without the danger of rejection caused by the presence of the sugar in cross-species procedures known as xenografts or xenotransplantation. ..."

FDA approves genetically altering pigs, potentially for drugs or transplants

A probiotic therapy for patients with inflammatory bowel disease

Good news!

"Now, a research team ... has developed a living-material approach that uses a strain of genetically engineered E.coli Nissle gut bacteria as a locally acting probiotic. The engineered bacteria produce a network of nanofibers that directly binds to mucus to fill inflamed areas like a patch, shielding them from gut microbes and environmental factors. ... “The Probiotic Associated Therapeutic Curli Hybrids (PATCH) approach, as we named it, creates a biocompatible, mucoadhesive coating that functions as a stable, self-regenerating Band-Aid and provides biological cues for mucosal healing.”"

A probiotic therapy for patients with inflammatory bowel disease – Harvard Gazette: A genetically programmed living hydrogel material that facilitates intestinal wound healing is being considered for development as a probiotic therapy for patients with inflammatory bowel disease.

Weizmann Institute develops CO2-consuming bacteria

Very beneficial for humanity that Jews/Israelis believe in only one God and it is not climate change! Jews believe in "Do not worship idols (e.g. climate change)!". Christians believe in the Apocalypse, Jews in the coming of the Messiah! Which one do you prefer?

This research most likely has also implications for artificial photosynthesis!

"Scientists at the Weizmann Institute have developed bacteria nourished only from carbon dioxide. The bacteria build their entire biomass from carbon in the air. ... The bacteria ... were weaned completely from sugar, and now subsist entirely on a diet of CO2, which they obtain from their surroundings, so that they live off the air and build their entire mass from carbon in the atmosphere. ... The researchers ... decided to try applying the no-sugar challenge to E. coli bacteria. First, they mapped the genes essential to the process of fixating carbon and attached several of them to the genome of the bacteria in their laboratory. In addition, in order to replace the role of solar energy in photosynthesis, they inserted a gene into the bacteria that enabled them to derive energy from an available material called formate. The scientists sequenced the genome of the bacteria weaned from sugar in order to identify the mutations that changed their nutritional habits. Unexpectedly, they found relatively few genetic changes - a few changes involving synchronization of the carbon fixation process, a few changes involving transcription - regulating how existing genes are turned on and off, and several more changes whose role was unclear"

"So, Ron Milo, a synthetic biologist at the Weizmann Institute of Science in Rehovot, Israel, and his colleagues decided to see whether they could transform E. coli [heterotroph] into an autotroph. ... hey inserted the gene for an enzyme that enabled the microbe to eat formate, one of the simplest carbon-containing compounds, and one other strains of E. coli can’t eat. The microbes could then transform the formate into ATP, an energy-rich molecule that cells can use. That diet gave the microbe the energy it needed to use the second batch of three new enzymes it received—all of which enabled it to convert CO2 into sugars and other organic molecules. The researchers also deleted several enzymes the bacterium normally uses for metabolism, forcing it to depend on the new diet to grow."

Weizmann Institute develops CO2-consuming bacteria - Globes: The bacteria could help reduce greenhouse gases in the atmosphere.

This microbe no longer needs to eat food to grow, thanks to a bit of genetic engineering

Soldiers Without Pain Or Fear

Posted: 8/23/2019

“A new approach to pain eradication mimics [a] mutation and has been demonstrated in lab mice. Startup Navega Therapeutics plans to develop a CRISPR treatment to block severe pain caused by diabetes, cancer, or car accidents ... The first tests of CRISPR gene therapies on humans began only recently, and it’s a huge gamble. Removing people’s ability to feel pain comes with some big risks

Super soldiers: The studies are likely to be closely followed by the military. In 2017 Vladmit Putin said genetic engineering could create soldiers who feel no pain or fear, which could be “scarier than a nuclear bomb.

A rare case: A young Pakistani street performer [who walked on hot coal] died in 2006 after someone dared him to jump from a roof. ... He had a rare genetic disorder which meant he never felt pain. He had a mutation which disabled the SCN9A gene, which pays a key part in transmitting pain to the brain.

” (S1; emphasis added)

If you add to this robots or autonomous fighter planes and drones or the proliferation of nuclear or biological weapons, you have a major paradigm shift in military strategy and warfare.

The world will only be a safer place once all the belligerent authoritarian regimes or dictatorships of the world are defeated or replaced by human dignity and individual freedom respecting forms of government! Eternal vigilance is the price of freedom and peace on earth!

Sources (S):

1. The next trick for CRISPR is gene-editing pain away A family of street performers could walk on coals. Here’s how the secret of why they felt no pain could benefit others. (8/22/19)