Super-sized deletions: improved transposon excision screens using a mus309 mutant background.
Over the past two decades, a large collection of transposable elements inserted at various locations in the Drosophila melanogaster genome has been assembled. These transposons are frequently utilized in imprecise excision screens to generate deletions in genes of interest. In general, these screens involve genetic manipulations to combine a non-autonomous transposon and the appropriate transposase in individual male or female flies. DNA double-strand breaks are created via transposase action in both somatic and germline cells of these individuals and inaccurate repair events are recovered in the progeny. Because deletion-prone repair of transposon-induced double-strand breaks is rare, these screens generally require a significant investment of time and resources. We recently reported that conducting imprecise excision screens in mus309 mutant flies, which lack the Drosophila ortholog of the Bloom Syndrome helicase, results in an increase in both the number and size of deletions recovered. Here, we provide additional information for Drosophila researchers wishing to utilize this technique. In addition, we discuss how the general principle behind this technique can be applied in other contexts where double-strand breaks are being generated for the purpose of genome modification.
Witsell
Alice
A
Department of Biology, Tufts University, Medford, MA, USA.
Kane
Daniel P
DP
McVey
Mitch
M
D000818:Animals; D053903:DNA Breaks, Double-Stranded; D004251:DNA Transposable Elements / Q000235:genetics*; D029721:Drosophila Proteins / Q000235:genetics*; D004331:Drosophila melanogaster / Q000235:genetics*; D005260:Female; D005820:Genetic Testing; D049750:Genome, Insect; D008297:Male; D009154:Mutation; D053484:RecQ Helicases / Q000235:genetics*; D017384:Sequence Deletion
D004251:DNA Transposable Elements; D029721:Drosophila Proteins; C097151:Bloom syndrome protein; D053484:RecQ Helicases