Single-Strand Annealing Plays a Major Role in Double-Strand DNA Break Repair following CRISPR-Cas9 Cleavage in Leishmania

CRISPR-Cas9 genome editing relies on an efficient double-strand DNA break (DSB) and repair. Contrary to mammalian cells, the protozoan parasite lacks the most efficient nonhomologous end-joining pathway and uses microhomology-mediated end joining (MMEJ) and, occasionally, homology-directed repair to...

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Veröffentlicht in:	mSphere 2019-08, Vol.4 (4)
Hauptverfasser:	Zhang, Wen-Wei, Matlashewski, Greg
Format:	Artikel
Sprache:	eng
Schlagworte:	Annealing Cell cycle Cell death CRISPR CRISPR-Associated Protein 9 - genetics CRISPR-Cas Systems Deoxyribonucleic acid DNA DNA Breaks, Double-Stranded DNA damage DNA polymerase DNA Repair DNA, Protozoan - metabolism DNA, Single-Stranded - chemistry DNA-directed DNA polymerase Double-strand break repair Efficiency Gene amplification Gene Editing - methods Gene targeting Genome editing Genome, Protozoan Genomes Homology Leishmania Leishmania - genetics Mammalian cells Miltefosine Molecular Biology and Physiology Mutation Parasitic diseases Protozoa Recombination, Genetic Staphylococcus aureus
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Zusammenfassung:	CRISPR-Cas9 genome editing relies on an efficient double-strand DNA break (DSB) and repair. Contrary to mammalian cells, the protozoan parasite lacks the most efficient nonhomologous end-joining pathway and uses microhomology-mediated end joining (MMEJ) and, occasionally, homology-directed repair to repair DSBs. Here, we reveal that predominantly uses single-strand annealing (SSA) (>90%) instead of MMEJ (
ISSN:	2379-5042 2379-5042
DOI:	10.1128/mSphere.00408-19