(
A) By injecting gRNAs only into Cas9-expressing embryos, we generated a deletion that encompassed the enitre
cav coding sequence (light blue rectangle). Below is a browser shot of the BLAT result (
https://genome.ucsc.edu/) using the PCR-amplified deleted region as the query. This allele is homozygous lethal (as are
cav[deletion] /
cav[1] flies). (
B) We generated a rescue trangene amplified from a 'wildtype' stock (w[1118]) and, using PhiC31 integrase, inserted the transgene on chromosome II. The transgene rescued viability of otherwise homozygous lethal cav[deletion] flies. (
C) Experimental evolution of two replicate populations of the
cav[deletion]/TM6b stock results in elevated HeT-A and TART genomic copy number, revealed by sequencing generation 0 and generation 10 populations. Y-axis is reads per million (rpm) at generation 10 minus rpm at generation 0 (with standard error). Circles represent the copy number estimates from the two replicate populations.