Researchers from Harvard’s Wyss Institute for Biologically Inspired Engineering have created a new gene editing tool that can enable researchers to perform millions of genetic experiments simultaneously. They call it the Retron Library Recombination (RLR) technique, and it uses segments of bacterial DNA called retrons that can produce fragments of single-stranded DNA.
When it comes to genre editing, CRISPR-Cas9 is probably the best known technique these days. It has created waves in the world of science over the last few years, giving scientists the tools they need to easily change DNA sequences. It is more accurate than previously used techniques and it has a wide range of potential applications, including life-saving treatments for various diseases.
However, the tool has some major limitations. It could be difficult to supply CRISPR-Cas9 materials in large numbers, which is still a problem for studies and experiments. The way the technique works can also be toxic to cells because the Cas9 enzyme ̵
CRISPR-Cas9 cuts physical DNA to incorporate the mutant sequence into its genome during the repair process. Meanwhile, retrons can introduce the mutant DNA strand into a replicating cell so that the strand can be incorporated into the DNA of the daughter cells. Furthermore, the sequences of retrons can serve as “barcodes” or “nameplates”, allowing researchers to track individuals in a bacterial pool. This means that they can be used for genome editing without damaging the native DNA, and they can be used to perform multiple experiments in a large mixture.
Wyss Institute researchers tested RLR on E coli bacteria and found that 90 percent of the population incorporated the retron sequence after making a few adjustments. They were also able to prove how useful it can be in massive genetic experiments. During their tests, they were able to find antibiotic resistance mutations in E coli by sequencing retron barcodes instead of sequencing individual mutants, making the process much faster.
The study’s co-author Max Schubert explained:
“RLR enabled us to do something impossible with CRISPR: we randomly chopped a bacterial genome, transformed these genetic fragments into single-stranded DNA in situ, and used them to screen millions of sequences simultaneously. RLR is a simpler , more flexible gene editing tool that can be used for highly multiplexed experiments, eliminating the toxicity often observed with CRISPR and improving researchers’ ability to explore mutations at the genome level …
For a long time, CRISPR was just considered a weird thing that bacteria did, and figuring out how to utilize it for genome engineering changed the world. Retrons is another bacterial innovation that may also provide some important advances. “
There is still work to be done before RLR can be used extensively, including improving and standardizing the editing speed. However, the team believes it can “lead to new, exciting and unexpected innovations.”
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