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NIH Research Festival

September 16 – 18, 2015

In vivo knockdown of RAD51 in mouse testicular cells reveals its role in meiotic homologous recombination in mammals

Friday, September 18, 2015 – Poster Session IV
12:00 – 1:30 p.m.

FAES Terrace

NIDDK

GEN-8

Authors

  • J Dai
  • O Voloshin
  • S Potapova
  • RD Camerini-Otero

Abstract

During meiosis, homologous recombination (HR) enables populations to adapt during evolution by producing new combinations of DNA sequences, and also ensures the stability of the karyotype of the organism by controlling the accurate segregation of homologous chromosome pairs. In most eukaryotes, recombinases RAD51 and DMC1 play an essential role in the repair of double strand breaks during meiotic HR. In mammals, DMC1 knockout results in an arrest of meiosis at early prophase I stage, without completion of HR. However, the function of RAD51 during meiotic HR in mammals remains unclear, due to the embryonic lethality of the RAD51 knockout mouse. Here we present our functional studies of RAD51 during mouse spermatogenesis using siRNA to knockdown RAD51 both in vivo by injecting siRNAs into mouse seminiferous tubules and in an in vitro spermatogenesis system with cultured spermatocytes. Our results reveals that RAD51 is indispensable for mouse meiotic HR, and that the knockdown of RAD51 leads to the quick death of spermatocytes from early prophase I stage through a P53 dependent apoptotic pathway. The RAD51 knockdown phenotype can be rescued by the recruitment of RAD51 wild type protein but not a RAD51 mutant protein. The system we have developed provides an experimental setup for the study of a number of meiotic proteins.

Scientific Focus Area: Genetics and Genomics

This page was last updated on Friday, March 26, 2021

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