Researchers of Wyss Institute for Biologically Inspired Engineering at Harvard University have reportedly created a novel gene-editing tool that could allow scientists to simultaneously conduct nearly millions of genetic experiments. The new gene-editing tool is named as the RLR (Retron Library Recombineering) method, it further utilizes segments of bacterial DNA named retrons which could produce single-stranded DNA fragments.
With regards to gene editing, CRISPR-Cas9 is perhaps the most prominent technique today. It has been making waves within the science world in the last few years, providing researchers the tool, they require to be able to easily modify DNA sequences. The tool is much more accurate than the earlier used methods, and it also has an extensive variety of potential applications, which includes life-saving treatments for numerous diseases.
But the tool has a few key limitations. It might be difficult to deliver CRISPR-Cas9-associated materials in huge numbers, which stays as an issue for experiments as well as studies for one. Further, the way the method works could be toxic to cells, since the Cas9 enzyme, the restriction enzymes in charge of cutting DNA strands, usually cuts non-target sites as well.
CRISPR-Cas9 technique cuts DNA to take in the genetic sequence into its genome mainly throughout the process of repair. In the meantime, the retrons could introduce the mutant DNA strand straight into a replicating cell, so that the strand could be mixed into the DNA of the daughter cells.
The scientists of the Wyss Institute tested Retron Library Recombineering on E. coli bacteria and discovered that nearly 90% of the overall population comprised the retron sequence after making some changes. They were further able to prove how beneficial it could be in large genetic experiments. The scientists were also able to discover antibiotic resistance mutations in E. coli bacteria by sequencing the barcodes of retrons rather than sequencing individual mutants, making the process much faster.
While, a lot of work still needs to be done before RLR could be widely utilized, including enhancing and standardizing its overall editing rate.