Researchers at the Harvard Institute for Biological Inspiration Wyss have created a new gene editing tool that can enable scientists to perform millions of genetic experiments simultaneously. They call it the Retron Library Recombination (RLR) technique and use segments of bacterial DNA called retrons, which can produce fragments of single-stranded DNA.
When it comes to gene editing, CRISPR-Cas9 is perhaps the most well-known technique today. Over the past few years, he has been making waves in the scientific world, giving researchers the tools they need to be able to easily change DNA sequences. It is more accurate than previously used techniques and has a wide variety of potential applications, including life-saving treatments for various diseases.
However, the tool has some major limitations. It can be difficult to deliver materials in CRISPR-Cas9 in large numbers, which remains a problem for research and experimentation for one. Also, the way the technique works can be toxic to cells, as the enzyme Cas9 ̵
CRISPR-Cas9 physically cleaves DNA to incorporate the mutant sequence into its genome during the repair process. In the meantime, the 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. In addition, retron sequences can serve as “barcodes” or “name tags”, allowing scientists to track individuals in a group of bacteria. This means that they can be used to edit the genome without damaging the natural DNA and can be used to perform many experiments in one large mixture.
Scientists from the Wyss Institute tested RLR on E. coli bacteria and found that 90 percent of the population included the retron sequence after performing several pinches. They have also been able to prove how useful it can be in massive genetic experiments. During the tests, they were able to detect mutations in antibiotic resistance E. coli by sequencing the barcodes of the retrons instead of the sequence of individual mutants, which makes the process much faster.
Study co-author Max Schubert explained:
“RLR has allowed us to do something that is impossible to do with CRISPR: we randomly cut a bacterial genome, turned these genetic fragments into single-stranded DNA in situ, and used them to screen millions of sequences at once. RLR is simpler, more a flexible gene editing tool that can be used for highly multiplexed experiments, which eliminates the toxicity often observed in CRISPR and improves the ability of researchers to study mutations at the genome level …
For a long time, CRISPR was simply considered a strange thing that bacteria do, and figuring out how to use it for genomic engineering changed the world. Retrons are another bacterial innovation that can also provide some important advances. “
Work still needs to be done before the RLR can be widely used, including improving and standardizing editing speed. However, the team believes it can “lead to new, exciting and unexpected innovations”.
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