Researchers at Harvard’s Wyss Institute for Biologically Inspired Engineering have created a new gene editing tool that can help scientists perform millions of genetic experiments simultaneously. They call it the Retron Library Recombineering (RLR) technique and use a portion of the bacterial DNA called reton, which can form a single-stranded piece of DNA.
When it comes to gene editing, CRISPR-Cas9 is probably the best-known technique today. In recent years it has made waves in the scientific world, giving researchers the tools necessary to easily modify DNA sequences. It is more accurate than previously used techniques and has a wide range of applications, including life-saving treatments for various ailments.
However, this tool has some important limitations. It can be difficult to deliver large volumes of CRISPR-Cas9 material, which remains a problem for studies and trials. In addition, the way this technique works can be cytotoxic because the enzyme Cas9 ̵1; the molecular “scissors” that cut the DNA strands – often cuts off non-target areas as well.
CRISPR-Cas9 cuts DNA to integrate the mutation sequence into its genome during the repair process. At the same time, retons are able to carry the mutated DNA strands into replicated cells so that the strands can integrate with the daughter cell DNA. The reton sequence can also function as a “barcode” or “name tag”, enabling scientists to track individuals in the bacterial population. That means it can be used for genome editing without damaging the original DNA, and it can be used to perform multiple experiments in a single large mixture.
Wyss Institute scientists have tested the RLR on E. coli Bacteria, and it was found that 90 percent of the population included the reton sequence after they made small adjustments. They can also prove useful in large-scale genetic experiments. During the tests, they were able to find antibiotic resistance mutations in E. coli By sequencing retrons’ barcodes rather than sequencing individual mutations, the process is much faster.
Max Schubert, the study’s first co-author, explains:
“RLR allows us to do something impossible with CRISPR: We randomly chop the bacterial genome, transform those genetic parts into single-stranded DNA in situ and use it to screen millions of sequences.” Simultaneously, the RLR is simpler and more. A flexible gene-editing tool that can be used for high-multiplexing experiments, eliminating the toxicities commonly encountered with CRISPR and improving the ability of researchers to explore genome-level mutations …
For a long time, CRISPR was the bizarre thing bacteria did, and figuring out how to control it for genome engineering changed the world. Retrons are another bacterial innovation that could make some important advances as well.
There is still work to be done before the RLR can be widely used, including improving and setting the correction rate standards. However, the work team believes that it can “Leading to new exciting and unexpected innovations”
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