Meiosis and homologous recombination (HR) are critical functions in the reproduction, evolution, and biology of higher eukaryotes. HR is needed for proper chromosome segregation during meiosis, and repair of DNA damage in somatic cells. This proposal centers on genetic control of meiotic recombination, meiosis, and repair of DNA by HR in the mouse. Two basic strategies will be taken to identify and characterize genes involved in meiosis and HR. One is a gene-driven approach to analyze the function of the RecA homologs Dmc1 and Rad51d in meiosis. A mutation of the meiosis-specific Dmc1 gene causes meiotic arrest from chromosome synapsis failure. To better define the underlying defect, the null allele will be combined with transgenes and mutations that perturb checkpoint function, double strand break (DSB) induction, and an alternative HR pathway. These tests are designed to bypass the meiotic arrest, and obtain gametes for analysis of HR and chromosome segregation. Since a null mutation of Rad51d causes embryonic lethality, we will create a conditional knockout to evaluate this gene's role in meiotic HR. The other strategy for identifying meiosis and HR genes exploits a novel technology for whole genome mutagenesis using ES cells. One mutant we generated with this approach, mei1, causes sterility from meiotic prophase arrest. It exhibits defective chromosome synapsis, and failure to load Rad51 onto meiotic chromosomes. We will positionally clone mei1, study its gene product, and also characterize the defects in a second sterile mutation, mei2. Screens for additional meiotic mutants will be performed. Finally, a screen for DSB repair mutations will be conducted directly in mutagenized ES cells, enabling the derivation of mutant mice without the extreme husbandry requirements of traditional mutagenesis.

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Research Project (R01)
Project #
3R01GM045415-11S1
Application #
6606779
Study Section
Special Emphasis Panel (ZRG1 (02))
Program Officer
Anderson, Richard A
Project Start
1992-02-01
Project End
2004-12-31
Budget Start
2002-01-01
Budget End
2002-12-31
Support Year
11
Fiscal Year
2002
Total Cost
$34,215
Indirect Cost
Name
Jackson Laboratory
Department
Type
DUNS #
042140483
City
Bar Harbor
State
ME
Country
United States
Zip Code
04609
Rinaldi, Vera D; Bolcun-Filas, Ewelina; Kogo, Hiroshi et al. (2017) The DNA Damage Checkpoint Eliminates Mouse Oocytes with Chromosome Synapsis Failure. Mol Cell 67:1026-1036.e2
McNairn, Adrian J; Rinaldi, Vera D; Schimenti, John C (2017) Repair of Meiotic DNA Breaks and Homolog Pairing in Mouse Meiosis Requires a Minichromosome Maintenance (MCM) Paralog. Genetics 205:529-537
Rinaldi, Vera D; Hsieh, Kristin; Munroe, Robert et al. (2017) Pharmacological Inhibition of the DNA Damage Checkpoint Prevents Radiation-Induced Oocyte Death. Genetics 206:1823-1828
Singh, Priti; Schimenti, John C (2015) The genetics of human infertility by functional interrogation of SNPs in mice. Proc Natl Acad Sci U S A 112:10431-6
Singh, Priti; Schimenti, John C; Bolcun-Filas, Ewelina (2015) A mouse geneticist's practical guide to CRISPR applications. Genetics 199:1-15
Luo, Yunhai; Hartford, Suzanne A; Zeng, Ruizhu et al. (2014) Hypersensitivity of primordial germ cells to compromised replication-associated DNA repair involves ATM-p53-p21 signaling. PLoS Genet 10:e1004471
Bolcun-Filas, Ewelina; Rinaldi, Vera D; White, Michelle E et al. (2014) Reversal of female infertility by Chk2 ablation reveals the oocyte DNA damage checkpoint pathway. Science 343:533-6
Handel, Mary Ann; Eppig, John J; Schimenti, John C (2014) Applying ""gold standards"" to in-vitro-derived germ cells. Cell 157:1257-61
Qiao, Huanyu; Prasada Rao, H B D; Yang, Ye et al. (2014) Antagonistic roles of ubiquitin ligase HEI10 and SUMO ligase RNF212 regulate meiotic recombination. Nat Genet 46:194-9
Schimenti, Kerry J; Feuer, Sky K; Griffin, Laurie B et al. (2013) AKAP9 is essential for spermatogenesis and sertoli cell maturation in mice. Genetics 194:447-57

Showing the most recent 10 out of 19 publications