On November 16th, published in a study on Nature (title: In vivo genome editing via CRISPR/Cas9 mediated homology-independent targeted integration), scientists at the Salk Institute discovered another "magic" of the genetic editing artifact CRISPR. . For the first time, researchers have confirmed that CRIPSR-based technology can insert DNA into targeted locations of non-dividing cells. In addition, the study also confirmed that this technology can partially restore the visual response of blind mice.
Professor Juan Carlos Izpisua Belmonte, the author of the paper, said: "We are very excited because this is something we could not do before. We entered the cell that could not be split for the first time and were able to modify the DNA arbitrarily. The possible application value of this discovery. It is very huge."
To date, DNA editing techniques, including the CRISPR-Cas9 system, have been most effective in dividing cells, such as cells in the skin or intestines. This new technology invented by the Salk Institute (Salk for short) is 10 times more efficient at inserting new DNA into dividing cells than other techniques. This will make this new technology a very promising tool in research and medicine. But more importantly, Salk technology has enabled scientists to successfully insert new genes into precise DNA positions in mature cells that no longer divide. These cells that no longer divide include the eyes, brain, pancreas or heart cells. This breakthrough offers new possibilities for therapeutic applications of these cells.
From left: Jun Wu, Reyna Hernandez-Benitez, Keiichiro Suzuki and Juan Carlos Izpisua Belmonte
How to achieve it?
To achieve this result, the scientists targeted a DNA repair cell pathway called non-homologous end-joining (NHEJ). The role of this pathway is to repair conventional DNA breaks. They combined this process with the existing gene editing technology CRISPR to successfully insert new DNA into precise locations of non-dividing cells.
Keiichiro Suzuki, co-first author of the paper, said: "The use of this NHEJ pathway to insert new DNA is a revolutionary result in the field of genome editing. No one has done this before."
Specifically, first, the research team set out to optimize the NHEJ mechanism for use with the CRISPR-Cas9 system. They created a custom insertion package consisting of a nucleic acid cocktail and called it homology-independent targeted integration (HITI). Subsequently, they used an inert virus to present HITI genetic instructions to the neurons.
Jun Wu, the co-first author of the paper, said: "This is the first sign that HITI may play a role in non-dividing cells." The team then successfully presented the structure into the brains of adult mice.
Pictured is a part of the adult mouse brain. Cell nuclei are blue and genome-edited neurons are green.
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