Scientists at Harvard University have invented a new gene editing technique that can introduce up to millions of mutated genes at once without harming the original DNA and greatly speeding up the process of finding mutated genes.
The commonly heard CRISPR editing technique uses a biological enzyme to cut into a specific part of DNA and replace the cut-out part with a new fragment while the DNA repairs itself. However, this technique is difficult to solve the problem of off-targeting, which will have unknowable consequences if it occurs, and in addition it is difficult to perform a large number of edits at the same time.
A new editing method invented by the Wyss Institute for Biologically Inspired Engineering at Harvard University, called Retron Library Recombineering (RLR), replicates a new fragment in a cell during The new method, called Retron Library Recombineering (RLR), introduces a new DNA fragment into a daughter cell at the time of replication, thus completing gene editing without harming the original DNA.
The study describes that the retrotransposon (Retron) can generate deformed versions of many millions of genes at the same time, so it is possible to experiment with many cells at the same time to create complex combinations of deformed versions of genes.
In addition, the newly introduced retrotransposon sequences act as “barcodes” or tags, helping scientists track down specific variants in a large number of mutated cells, greatly reducing the search process.
Researchers conducted editing experiments with Escherichia coli (E. coli) and showed that 90% of the bacteria incorporated the edited retrotransposon sequence. Previously, in order to find the genes in which drug resistance variants appeared, researchers needed to sequence them one by one. With the RLR method, the researchers were able to find the genes with drug-resistant variants directly through a “barcode” screen of sequenced retrotransposons, dramatically increasing the speed of the search.
One of the researchers, Max Schubert, said, “CRISPR was seen as a weird thing that bacteria did for a long time before it was discovered that it could control bacterial CRISPR behavior for useful gene editing. Reverse transcripts are yet another innovative technology relevant to bacteria that will hopefully lead to important developments.”
The study was published May 4 in the Proceedings of the National Academy of Sciences (PNAS).
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