Abstract
Meiotic homologous recombination (HR) is important for proper chromosomal segregation during gametogenesis and facilitates evolutionary adaptation via genomic reshuffling. In most eukaryotes, HR is mediated by two recombinases, the ubiquitous RAD51 and the meiosis-specific DMC1. The role of RAD51 in mammalian meiosis is unclear and study of its function is limited due to embryonic lethality of RAD51 knockouts. Here, we developed an invivo meiotic knockdown and protein complementation system to study RAD51 during mouse spermatogenesis. We show that RAD51 is crucial during meiotic prophase and its loss leads to depletion of late prophase I spermatocytes through a p53-dependent apoptotic pathway. This phenotype is distinct from that observed in the DMC1 knockdown. Our meiotic knockdown and complementation system establishes an experimental platform for mechanistic studies of meiotic proteins with unknown functions or essential genes for which a testis-specific knockout is not possible.
Highlights
Meiotic homologous recombination (HR) is indispensable for generating new combinations of DNA sequences to enable populations to adapt during the course of evolution
Our results indicate that RAD51 plays an essential role in the progression of mammalian meiosis and it provides a framework for further molecular studies on RAD51 in mammalian meiotic HR
Depletion of DMC1 in Mouse Testis with an In Vivo Knockdown Approach To determine the role of RAD51 in mammalian meiotic HR, we developed an in vivo knockdown system based on the microinjection of small interfering RNAs (siRNAs) into seminiferous tubules of mouse testis
Summary
Dai et al develop an in vivo meiotic gene knockdown and protein complementation system to show that RAD51 has an essential role in mammalian meiosis. RAD51 loss in mouse testis leads to the depletion of late prophase I spermatocytes through a p53dependent pathway.
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