Use of a yeast site-specific recombinase to produce female germline chimeras in Drosophila.

TLDR

The authors develop the FLP‑DFS technique to efficiently generate female germline mosaics via heat‑shock‑induced FLP recombinase–mediated mitotic recombination, enabling analysis of maternal effects of X‑linked zygotic lethal mutations. FLP‑DFS triggers site‑specific homologous mitotic recombination in ovoD1 heterozygous flies using heat‑shock‑induced FLP recombinase, producing germline clones that are the sole cells capable of egg laying and whose parameters are optimized for efficient mosaic generation. The FLP‑DFS method recovers germline clones in over 90% of females, does not impair viability, and yields clones indistinguishable from those generated by X‑ray–induced mitotic recombination.

Abstract

Abstract We describe an efficient method for generating female germline mosaics by inducing site-specific homologous mitotic recombination with a yeast recombinase (FLP) which is driven by a heat shock promoter. These germline mosaics are produced in flies heterozygous for the agametic, germline-dependent, dominant female sterile (DFS) mutation ovoD1, where only flies possessing germline clones are able to lay eggs. This method, the "FLP-DFS" technique, is very efficient because more than 90% of females with germline clones can be recovered. We show that this heat-inducible, site-specific mitotic recombination system does not affect viability and that the germline clones recovered are physiologically the same as those created by X-ray induced mitotic recombination. We describe the parameters of FLP-recombinase induced germline mitotic recombination and the use of the "FLP-DFS" technique to analyze the maternal effect of X-linked zygotic lethal mutations.