title: Transcription-replication collision increases recombination efficiency between plasmids.
abstract: It has been proposed that the stalling of the replication forks can induce homologous recombination in several organisms, and that arrested replication forks may offer nuclease targets, thereby providing a substrate for proteins involved in double-strand repair. In this article, we constructed a plasmid with the potential for transcription-replication collision (TRC), in which DNA replication and RNA transcription occur on the same DNA template simultaneously. Theoretically, transcription will impede DNA replication and increase homologous recombination. To validate this hypothesis, another plasmid was constructed that contained a homologous sequence with the exception of some mutated sites. Co-transfection of these two plasmids into 293T cells resulted in increased recombination frequency. The ratio of these two plasmids also affected the recombination frequency. Moreover, we found high expression levels of RAD51, which indicated that the increase in the recombination rate was probably via the homologous recombination pathway. These results indicate that mutant genes in plasmids can be repaired by TRC-induced recombination.
authors.affiliation: National Laboratory of Medical Molecular Biology, Institute of Basic Medical Sciences, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing 100005, China; Guangzhou Fuda Cancer Hospital, No. 2, Tangdexi Road, Tang Xia, Tianhe District, Guangzhou City, Guangdong Province 510665, China.
authors.forename: Li
authors.initials: L
authors.lastname: Jialiang
authors.forename: Chen
authors.initials: C
authors.lastname: Feng
authors.forename: Xu
authors.initials: X
authors.lastname: Zhen
authors.forename: Chen
authors.initials: C
authors.lastname: Jibing
authors.forename: Lv
authors.initials: L
authors.lastname: Xiang
authors.forename: Zhang
authors.initials: Z
authors.lastname: Lingling
authors.forename: Liu
authors.initials: L
authors.lastname: Depei
mesh_terms: D004247:DNA / Q000235:genetics* / Q000378:metabolism; D004260:DNA Repair; D004261:DNA Replication; D015870:Gene Expression; D057809:HEK293 Cells; D059765:Homologous Recombination; D006801:Humans; D009154:Mutation; D010957:Plasmids; D051135:Rad51 Recombinase / Q000235:genetics / Q000378:metabolism; D014158:Transcription, Genetic; D014170:Transformation, Genetic
chemical_list: D004247:DNA; D051135:Rad51 Recombinase
keywords: CMV; EGFP; FCM; Homologous recombination; Recombination frequency; Replication; TRC; Transcription; Transcription–replication collision; cytomegalovirus; enhanced green fluorescent protein; flow cytometry; transcription–replication collision