Researchers analysed thousands of laboratory-made plasmids and discovered that nearly half of them had defects, raising questions of experimental reproducibility.
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A new gene-editing tool has enabled Harvard's Wyss Institute for Biologically Inspired Engineering scientists to accomplish a feat that wouldn't be possible to do with CRISPR, according to a press release.
The group of researchers created what they call the “Retron Library Recombineering” (RLR) technique, which could allow scientists to run millions of genetic experiments at the same time.
This tool, described in a recent paper in PNAS, employs retrons, which are bacterial DNA segments that undergo reverse transcription to generate single-stranded DNA fragments (ssDNA). RLR produces up to millions of mutations concurrently in bacterial cells and “barcodes” mutant cells, enabling the whole pool to be screened at once. This way large quantities of data can be quickly produced and analyzed.
But why is this important? Well, because it overcomes the major limitations of CRISPR-Cas9, a groundbreaking technology that can be used to edit genes. Overall, it is difficult for scientists to deliver CRISPR-Cas9 materials in large numbers, and it can sometimes be toxic to cells since the Cas9 enzyme, the molecular "scissors" that cut strands of DNA, often cuts unintended...