Gene editing with CRISPR can cause off-target mutations, but this seems to happen less often with an enzyme that cuts one of the strands of DNA instead of both
at the University of California, San Diego, and their colleagues have developed a new form of CRISPR that can more efficiently insert correct DNA sequences at the site of a mutation, with fewer off-target effects.
The method uses a variant of the Cas9 enzyme called a nickase, which only cuts one strand of the DNA double helix. “We found that ‘softly’ nicking, or cutting, one strand of the DNA is even more efficient than making a clean double-stranded break,” says Bier.The researchers tested the approach in fruit flies that had a mutation that turned their eyes white instead of red.
The team didn’t introduce any extra pieces of DNA as a template for the cell to correct the mutation on the chromosome, so the molecular machinery must have used the other chromosome – inherited from the other parent – as a template. The team was able to confirm this was the case. “There’s accumulating evidence that when you create damage to one chromosome in a mammalian cell, then that somehow recruits the other chromosome. Then the region that’s broken gets the Band-Aid from the other chromosome,” says Bier.
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