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Towards an understanding of C9orf82 protein/CAAP1 function
Muhammad Assad Aslam1, Mir Farshid Alemdehy1, Colin E J Pritchard2
1Division of Tumor Biology & Immunology, The Netherlands Cancer Institute, Amsterdam, The Netherlands.
Plos One
|January 11, 2019
Summary
The C9orf82 protein
Area of Science:
- Cellular biology
- Molecular genetics
- Apoptosis research
Background:
- C9orf82 protein (also known as CAAP1) has been linked to apoptosis regulation and drug resistance.
- Previous studies suggested C9orf82 delays DNA double-strand break (DSB) repair, sensitizing cells to apoptosis.
- Its precise function in vivo remained unclear.
Purpose of the Study:
- To investigate the in vivo and ex vivo function of C9orf82.
- To generate a C9orf82 knockout mouse model using CRISPR/Cas9 technology.
- To assess C9orf82's role in DNA repair and apoptosis.
Main Methods:
- CRISPR/Cas9 gene editing in mouse zygotes to create C9orf82 knockout (C9orf82ko/ko) mice.
- Phenotypic analysis of C9orf82ko/ko mice, including macroscopic, histological, and lymphocyte development assessments.
- Ex vivo studies using primary pre-B cell cultures and mouse embryo fibroblasts (MEFs) to evaluate sensitivity to DNA damage and repair kinetics (γH2AX focus formation).
Main Results:
- C9orf82ko/ko mice were viable and showed no overt abnormalities.
- Lymphocyte development and class switch recombination (CSR) were normal in C9orf82-deficient mice.
- C9orf82ko/ko and wild-type cells exhibited comparable sensitivity to DNA damaging agents (etoposide, doxorubicin) and similar DNA repair kinetics.
Conclusions:
- C9orf82 protein does not appear to play a significant role in delaying the repair of Topoisomerase II-induced DSBs.
- The study challenges the previously proposed function of C9orf82 in regulating apoptosis through modulation of DSB repair.
- The generated C9orf82 knockout models provide valuable tools for future research into the protein's actual function.
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