Characterization and regulation of cell cycle-independent noncanonical gene targeting.
Nat Commun
; 15(1): 5044, 2024 Jun 18.
Article
em En
| MEDLINE
| ID: mdl-38890315
ABSTRACT
Homology-dependent targeted DNA integration, generally referred to as gene targeting, provides a powerful tool for precise genome modification; however, its fundamental mechanisms remain poorly understood in human cells. Here we reveal a noncanonical gene targeting mechanism that does not rely on the homologous recombination (HR) protein Rad51. This mechanism is suppressed by Rad52 inhibition, suggesting the involvement of single-strand annealing (SSA). The SSA-mediated gene targeting becomes prominent when DSB repair by HR or end-joining pathways is defective and does not require isogenic DNA, permitting 5% sequence divergence. Intriguingly, loss of Msh2, loss of BLM, and induction of a target-site DNA break all significantly and synergistically enhance SSA-mediated targeted integration. Most notably, SSA-mediated integration is cell cycle-independent, occurring in the G1 phase as well. Our findings provide unequivocal evidence for Rad51-independent targeted integration and unveil multiple mechanisms to regulate SSA-mediated targeted as well as random integration.
Texto completo:
1
Base de dados:
MEDLINE
Assunto principal:
Ciclo Celular
/
Marcação de Genes
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Proteína 2 Homóloga a MutS
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Rad51 Recombinase
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Proteína Rad52 de Recombinação e Reparo de DNA
Idioma:
En
Ano de publicação:
2024
Tipo de documento:
Article