Bisulfite treatment and single-molecule real-time sequencing reveal D-loop length, position, and distribution.

ABSTRACT: Displacement loops (D-loops) are signature intermediates formed during homologous recombination. Numerous factors regulate D-loop formation and disruption, thereby influencing crucial aspects of DNA repair, including donor choice and the possibility of crossover outcome. While D-loop detection methods exist, it is currently unfeasible to assess the relationship between D-loop editors and D-loop characteristics such as length and position. Here, we developed a novel in vitro assay to characterize the length and position of individual D-loops with near base-pair resolution and deep coverage, while also revealing their distribution in a population. Non-denaturing bisulfite treatment modifies the cytosines on the displaced strand of the D-loop to uracil, leaving a permanent signature for the d

SUBMITTER: Shah SS 

PROVIDER: S-EPMC7695462 | biostudies-literature | 2020 Nov

REPOSITORIES: biostudies-literature