Multiplexed conditional genome editing with Cas12a in <i>Drosophila</i>
Fillip Port, Maja Starostecka, Michael Boutros
Abstract
Significance CRISPR-Cas genome engineering has enabled scientists to modify the genome of the popular model organism Drosophila melanogaster with unprecedented ease. However, so far only the CRISPR nuclease Cas9 has been used for this task. Additional nucleases could expand the genomic target space and facilitate modification of independent cell populations in the same organism. Here we describe genome editing with Cas12a in Drosophila . Cas12a recognizes target sites orthogonal to Cas9, can use compact arrays of crRNAs for multiplexed gene editing, and conditional expression leads to tissue-specific mutagenesis. We also show that a point mutant in Cas12a substantially increases editing rates. This work significantly expands the CRISPR toolbox in Drosophila and will inform the development of similar systems in other organisms.