MIT New Medical Research: When the CRISPR Gene Editor Meets Parkinson
Release date: 2017-10-17
Based on an improved version of the CRISPR gene editing system, researchers at the Massachusetts Institute of Technology have developed a new method to screen for genes that protect specific diseases. The study was funded by the Ellison Medical Foundation and the National Institutes of Health (NIH).
The data show that CRISPR (Clustered regular interspaced short palindromic repeats) is an immune weapon produced by the struggle between bacteria and viruses in the history of life evolution. Scientists will cut off the DNA of various target cells by operating the protein Cas9. This technology is called the CRISPR/Cas9 gene editing system and quickly became the hottest technology in life sciences.
CRISPR is often used to edit or delete genes in living cells. However, the MIT team applied it to a large number of randomly-opened genetic cell populations, enabling researchers to identify protein genes that protect cells associated with Parkinson's disease.
Timothy Lu, an associate professor of electrical engineering and computer science and bioengineering at the Massachusetts Institute of Technology (MIT), said the new technology was published in the journal Molecular Cell. Not limited to Parkinson's disease, it also provides a new way to find drug targets for many diseases.
“The artistry of this technique is that it “targets†two or three genes for subsequent observations. But we believe that the genomes that need to be regulated to fit the disease are actually more than two or three genes. "Lu said. Lu is the core author of the study, and the authors include postdoctoral Chen Yingzhou and graduate student Fahim Farzadfard.
Is genetic editing "on or off"?
At the end of 2016, researchers from the University of Massachusetts Medical School and the University of Toronto in Canada found the first known CRISPR/Cas9 active “off switch†to provide better control for the CRISPR/Cas9 editor.
The CRISPR gene editing system consists of a DNA cleavage enzyme called Cas9 and a short RNA guide chain targeting a specific genomic sequence that "tells" where Cas9 should be "cut". Through this system, scientists can make targeted mutations in the genome of living animals, or delete genes, or insert new genes.
In this new study, the MIT research team inactivates the cleavage ability of Cas9 and designs a protein that, when combined with the target site, triggers transcription factors (meaning activation is required) Gene protein).
By transferring this version of Cas9 and the guide RNA strand into a single cell, the researcher can target a single gene sequence in a single cell. Depending on the specific leader sequence, each leader RNA may hit a single gene or multiple genes, allowing researchers to randomly screen for survival genes affecting cells throughout the genome.
"We take an objective and neutral approach, rather than selecting specific genes based on interest bias," Lu said. "In this way, we can screen out RNAs that have unusually strong protective activities in a neurodegenerative disease model."
The researchers used this technique in yeast cell alpha-synuclein, a genetically engineered protein that produces a drug associated with Parkinson's disease that forms in the brains of patients with Parkinson's disease. Bulk, usually toxic to yeast cells.
Based on this, the MIT team found a strong-acting RNA strand that makes cell activity much more efficient than other single genes that protect this yeast cell.
Further genetic screening revealed that many of the genes that are turned on by this RNA strand are molecular chaperone proteins that help other proteins "fold" into the correct shape. The researchers speculate that these chaperone proteins help to synthesize alpha-synuclein, which prevents it from forming clumps.
Other genes activated by the above RNA encode mitochondrial proteins that help cells regulate their energy metabolism, participate in packaging and transport proteins contained in other proteins. Now, researchers are studying whether these RNAs can activate these genes independently, or whether it activates one or more regulatory genes and activates other genes.
Protective effects
Lu said that once the researchers identified the genes in the yeast, they tested the analogs in human neurons in a laboratory dish, which also caused an excess of alpha-synuclein. On the other hand, these human genes also protect against alpha-synuclein-induced death, which can be tested as a method of treating Parkinson's disease.
Wilson Wong, assistant professor of biomedical engineering at Boston University, said the study highlights the diversity of practical applications of CRISPR/Cas9. "What's more interesting is that they can use yeast as a primer for gene screening and identify the toxicity of alpha-synuclein in mammalian cells to protect RNA," Wong, who did not participate in the study, said The study paves the way for the use of random RNA and yeast in complex human biology."
Lu's lab is now using this method to screen for genes associated with other diseases. Fortunately, researchers seem to have discovered genes that prevent aging.
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