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Sexual Healing: Examining the Role of Meiosis Genes in DNA Damage Repair
Author(s) -
Jasso Erika,
Hattabaugh Lettie,
Schurko Andrew
Publication year - 2015
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.29.1_supplement.879.5
Subject(s) - rad51 , biology , rna interference , dna repair , meiosis , gene , homologous recombination , genetics , gene silencing , dna damage , dna , microbiology and biotechnology , rna
Bdelloid rotifers are a class of microinvertebrates that have long lost the ability to reproduce sexually. Although bdelloids lack meiosis, their genomes contain genes ( SPO11 , MSH4 , MSH5 , and HOP1 ) that are specific to meiosis in eukaryotes. Bdelloids also have a remarkable DNA repair system that allows them to recover from damage induced by ionizing radiation. Our objective is to test the hypothesis that “meiotic” genes in bdelloids are maintained for a non‐meiotic function, specifically DNA repair. To address this, we used real‐time PCR to quantify meiotic gene expression in the bdelloid Adineta vaga after exposure to 280 Gray of ionizing radiation. DNA repair genes (such as MRE11 and RAD50 ) were upregulated (relative to non‐irradiated rotifers), but expression levels of meiotic genes remained constant. Since the absence of upregulation does not rule out a role for meiotic genes in DNA repair, we used RNA interference (RNAi) to investigate meiotic gene function. We explored two methods of inducing RNAi. For the feeding method, induction of RNAi was done by feeding bdelloids E. coli that produces double stranded RNA (dsRNA) copies of the target gene. For the soaking method, RNAi was induced by transfecting A. vaga with dsRNA copies of the target gene. For each method, real‐time PCR and western blots assessed gene knockdowns of HSP82 (heat‐shock protein) and RAD51 (recombination/DNA repair protein). The RNAi soaking method yielded the most reliable knockdowns, and it is being used to identify defects in DNA repair when the meiotic genes are knocked down. This will provide insight into the role of meiotic genes in bdelloid DNA repair. This work was supported by grants from the NCRR (5P20RR016460) & NIGMS (8P20GM103429) from the NIH.