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Salk scientists modify CRISPR to epigenetically treat diabetes, kidney disease, muscular dystrophy

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LA JOLLA—Salk scientists have created a new version of the CRISPR/Cas9 genome editing technology that allows them to activate genes without creating breaks in the DNA, potentially circumventing a major hurdle to using gene editing technologies to treat human diseases.

Most CRISPR/Cas9 systems work by creating “double-strand breaks” (DSBs) in regions of the genome targeted for editing or for deletion, but many researchers are opposed to creating such breaks in the DNA of living humans. As a proof of concept, the Salk group used their new approach to treat several diseases, including diabetes, acute kidney disease, and muscular dystrophy, in mouse models.

“Although many studies have demonstrated that CRISPR/Cas9 can be applied as a powerful tool for gene therapy, there are growing concerns regarding unwanted mutations generated by the double-strand breaks through this technology,” says Juan Carlos Izpisua Belmonte, a professor in Salk’s Gene Expression Laboratory and senior author of the new paper, published in Cell on December 7, 2017. “We were able to get around that concern.”

Affecting Up to 216,000 Studies — Popular Genetic Method Found To Be Deeply Flawed

According to recent research from Sweden’s Lund University, the most commonly used analytical method in population genetics is deeply flawed. This could have caused incorrect results and misconceptions regarding ethnicity and genetic relationships. The method has been used in hundreds of thousands of studies, influencing findings in medical genetics and even commercial ancestry tests. The findings were recently published in the journal Scientific Reports.

The pace at which scientific data can be gathered is increasing rapidly, resulting in huge and very complex databases, which has been nicknamed the “Big Data revolution.” Researchers employ statistical techniques to condense and simplify the data while maintaining the majority of the important information in order to make the data more manageable. PCA (principal component analysis) is perhaps the most widely used approach. Imagine PCA as an oven with flour, sugar, and eggs serving as the input data. The oven may always perform the same thing, but the ultimate result, a cake, is highly dependent on the ratios of the ingredients and how they are mixed.

“It is expected that this method will give correct results because it is so frequently used. But it is neither a guarantee of reliability nor produces statistically robust conclusions,” says Dr. Eran Elhaik, Associate Professor in molecular cell biology at Lund University.

A genetically encoded tool to increase cellular NADH/NAD+ ratio in living cells

Impaired reduction/oxidation (redox) metabolism is a key contributor to the etiology of many diseases, including primary mitochondrial disorders, cancer, neurodegeneration, and aging. However, mechanistic studies of redox imbalance remain challenging due to limited strategies which can perturb cellular redox metabolism and model pathology in various cellular, tissue, or organismal backgrounds without creating additional and potentially confounding metabolic perturbations. To date, most studies involving impaired redox metabolism have focused on oxidative stress and reactive oxygen species (ROS) production; consequently, less is known about the settings where there is an overabundance of reducing equivalents, termed reductive stress. NADH reductive stress has been modeled using pharmacologic inhibition of the electron transport chain (ETC) and ethanol supplementation. Still, both these methods have significant drawbacks. Here, we introduce a soluble transhydrogenase from E. coli (Ec STH) as a novel genetically encoded tool to promote NADH overproduction in living cells. When expressed in mammalian cells, Ec STH, and a mitochondrially-targeted version (mito Ec STH), can elevate the NADH/NAD+ ratio in a compartment-specific manner. Using this tool, we determine the metabolic and transcriptomic signatures of NADH reductive stress in mammalian cells. We also find that cellular responses to NADH reductive stress, including blunted proliferation, are dependent on cellular background and identify the metabolic reactions that sense changes in the cellular NADH/NAD+ balance. Collectively, our novel genetically encoded tool represents an orthogonal strategy to perturb redox metabolism and characterize the impact on normal physiology and disease states.

The authors have declared no competing interest.

Researchers identify potential gene marker for treating pancreatic cancer

Researchers at Mayo Clinic Comprehensive Cancer Center have identified a gene marker that may lead to a more effective, precision treatment for pancreatic ductal adenocarcinoma (PDAC). The researcher’s findings are published in Nature Cancer.

“Pancreatic ductal adenocarcinoma is one of the most lethal cancers,” says the paper’s senior author Zhenkun Lou, Ph.D. Dr. Lou says while Poly-ADP-ribose-polymerase inhibitors (PARPi) are now an FDA-approved option for standard maintenance therapy for patients with metastatic PDAC who harbor pathogenic germline BRCA1/2 mutations, only about 10 percent of patients with PDAC harbor pathogenic mutations of the homologous recombination (HR) genes. “This leaves most patients missing out on this encouraging treatment strategy,” says Dr. Lou.

In this study, Dr. Lou and his colleagues found that the protein METTL16 may be a new biomarker for PARPi treatment, and that PDAC with elevated expression of METTL16, may benefit from PARPi treatment.

Molecular Changes in the Brain in the Aftermath of a Traumatic Event May Help Explain Long-Term Susceptibility or Resilience

Summary: In mice genetically more susceptible to PTSD following a stressful event, researchers found an increased expression of cortisol receptors on neurons in the CA1 region of the dorsal hippocampus. Those increased receptors enabled an elevated expression of the HCN1 protein and TRIP8b, reducing neural excitability.

Source: medical college of georgia at augusta university.

Social avoidance is a common symptom of PTSD, and scientists working to better understand why have laboratory evidence that while stress hormone levels consistently increase in the immediate aftermath of a traumatic event, there can be polar opposite consequences in parts of the brain down the line.

Scientists Use CRISPR to Condense a Million Years of Evolution Into Mere Months

Chromosome-level engineering is a completely different beast: it’s like rearranging multiple paragraphs or shifting complete sections of an article and simultaneously hoping the changes add capabilities that can be passed onto the next generation.

Reprogramming life isn’t easy. Xiao Zhu’s DNA makeup is built from genetic letters already optimized by eons of evolutionary pressure. It’s no surprise that tinkering with an established genomic book often results in life that’s not viable. So far, only yeast have survived the rejiggering of their chromosomes.

The new study, published in Science, made the technology possible for mice. The team artificially fused together chunks from mice chromosomes. One fused pair made from chromosomes four and five was able to support embryos that developed into healthy—if somewhat strangely behaved—mice. Remarkably, even with this tectonic shift to their normal genetics, the mice could reproduce and pass on their engineered genetic quirks to a second generation of offspring.

Pigs With Gene Defect Provide New Perspectives for the Treatment of Alzheimer’s

Summary: New research in cloned pigs with a mutation of the SORL1 sheds light on Alzheimer’s development. The findings could pave the way for new treatments for the neurodegenerative disorder.

Source: Aarhus University.

For decades, researchers from all over the world have been working hard to understand Alzheimer’s disease. Now, a collaboration between the Department of Biomedicine and the Department of Clinical Medicine at Aarhus University has resulted in a flock of minipigs that could lead to a major step forward in the research and treatment of Alzheimer’s.

Genetic Divergence & Civilization

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As humanity reaches out to the stars and make new homes on strange new worlds, how will our genetics & DNA change under those alien planets?

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Episode 236; April 30, 2020

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Disordered Systems Mimic Genetic Evolution

Modern medicine forces bacteria to adapt: in response to antibiotic treatment, they either increase their fitness or die out. Whether a bacterial population survives or not depends on a combination of its genetics and environment—the antibiotic concentration—at a given moment. Now Suman Das of the University of Cologne, Germany, and colleagues simulate the effect on adaptation of an environment that is constantly changing [1]. Using a model that describes how slow-moving disordered systems respond to external forces, the researchers find that microbe evolution in changing drug concentrations exhibits hysteresis and memory formation. They use analytical methods and numerical simulations to connect these statistical physics concepts to bacterial drug resistance.

The team’s model examines changes in a bacterial population’s genetic sequences. By combining data on bacterial growth rates with statistical tools, the researchers describe how the bacterial genome can store information about both present and past drug concentrations. Their simulations start with a genetic sequence optimized for a certain antibiotic concentration. They then track how the sequence mutates when the concentration shifts to another value. When the concentration increases and then reduces to a lower value, the genetic route taken on the downward path depends on the changes on the upward path. How different the mutation routes are depends on the rate of concentration change.

The researchers find that this behavior mimics that of disordered systems driven by external forces, such as ferromagnetic materials subjected to magnetic fields or amorphous materials subjected to a shearing force. They say that while their approach focuses on the evolution of drug resistance, the framework can be adapted to other problems in evolutionary biology that involve changing environmental parameters.

New method allows scientists to determine all the molecules present in the lysosomes of mice

Small but mighty, lysosomes play a surprisingly important role in cells despite their diminutive size. Making up only 1–3% of the cell by volume, these small sacs are the cell’s recycling centers, home to enzymes that break down unneeded molecules into small pieces that can then be reassembled to form new ones. Lysosomal dysfunction can lead to a variety of neurodegenerative or other diseases, but without ways to better study the inner contents of lysosomes, the exact molecules involved in diseases—and therefore new drugs to target them—remain elusive.

A new method, reported in Nature on Sept. 21, allows scientists to determine all the molecules present in the lysosomes of any cell in mice. Studying the contents of these molecular recycling centers could help researchers learn how the improper degradation of cellular materials leads to certain diseases. Led by Stanford University’s Monther Abu-Remaileh, institute scholar at Sarafan ChEM-H, the study’s team also learned more about the cause for a currently untreatable neurodegenerative known as Batten disease, information that could lead to new therapies.

“Lysosomes are fascinating both fundamentally and clinically: they supply the rest of the cell with nutrients, but we don’t always know how and when they supply them, and they are the places where many diseases, especially those that affect the brain, start,” said Abu-Remaileh, who is an assistant professor of chemical engineering and of genetics.

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