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Using Next Generation Sequencing to Identify Mutations Associated with Repair of a CAS9-induced Double Strand Break Near the CD4 Promoter
Published on: March 31, 2022
DNA repair efficiency and thermotolerance in Drosophila melanogaster from "Evolution Canyon"
Achsa Lupu1, Antonina Pechkovskaya, Eugenia Rashkovetsky
1Institute of Evolution, University of Haifa, Mount Carmel, Haifa 31905, Israel.
Abstract:
The repair efficiency of four thermotolerant and four thermosensitive isofemale lines of Drosophila melanogaster originating from "Evolution Canyon" (Mt Carmel, Israel) was tested using 2-acetylaminofluorene (2-AAF) as mutagen. First, males of the standard laboratory line Canton S were treated with either 2-AAF solution or control solution. Then, females of the "Evolution Canyon" lines were crossed with treated (2-AAF or control solution) males and maintained at either 24 or 29 degrees C. Arbitrary primed PCR fingerprinting was employed as a method for genomic damage analysis in the resulting progeny (by scoring the frequency of lost DNA bands in F(1) progeny). Thermosensitive lines displayed significantly higher rates of change in the DNA fingerprint pattern after mutagenic presyngamic treatment followed by development at both temperatures, as well as after development under high temperature with no prior mutagenic treatment. The thermotolerant lines tended to show a lower level of mutation at both temperatures and after both treatments. One isofemale line showed a higher level of mutation at room temperature compared with increased temperature, after both control and mutagen treatment. The results suggest the existence of a relationship between DNA repair efficiency and thermotolerance, with thermotolerant lines tending to repair DNA more efficiently than thermosensitive ones.
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