Like other Drosophila researchers we’ve talked to, we are excited by these techniques and we can’t wait to try them out in the lab (Our new postdoc, Byoungchun Lee will be doing a lot of the work here) – CRISPR/Cas9-mediated mutagenesis will very likely develop into a “must-have” genetic technique for model organism labs.
Questions/comments we had: We mostly talked about how we would use this technique for our lab, with our favorite genes. In the Bassett et al paper the efficiency and frequency of mutagenesis for y and w looked good (although there was a high level of lethality – even in the non-injected controls ?!). But will this high efficiency hold true for all genes? Will genomic/chromatin context matter? (some of our favorite genes are in heterochromatic regions) Also, what’s the most efficient fly injecting/crossing scheme to generate stable mutant stocks (especially if the mutation efficiency ends up being quite low for some genes)? Finally, we’re interested to see how well the combination of CRISPR/Cas9 with donor oligos can be used to engineer specific alterations in our favourite genes. Gratz et al did a great job of using the approach to introduce attP sites into yellow, and presumably a similar approach can be used to introduce specific nucleotide substitutions into our favourite genes (e.g. to generate serine-to-alanine mutations at known phosphorylation sites).
Overall, two great papers. Exciting times ahead for fly genetics!