Oct 30, 2018

Mapping native R-loops genome-wide using a targeted nuclease approach

BioRxiv : the Preprint Server for Biology
Qingqing YanKavitha Sarma

Abstract

R-loops are three-stranded DNA:RNA hybrids that are pervasive in the eukaryotic and prokaryotic genomes and have been implicated in a variety of nuclear processes, including transcription, replication, DNA repair, and chromosome segregation. While R-loops may have physiological roles, the formation of stable, aberrant R-loops has been observed in disease, particularly neurological disorders and cancer. Despite the importance of these structures, methods to assess their distribution in the genome invariably rely on affinity purification, which requires large amounts of input material, is plagued by high level of noise, and is poorly suited to capture dynamic and unstable R-loops. Here, we present a new method that leverages the affinity of RNase H for DNA:RNA hybrids to target micrococcal nuclease to genomic sites that contain R-loops, which are subsequently cleaved, released, and sequenced. Our R-loop mapping method, MapR, is as specific as existing techniques, less prone to recover non-specific repetitive sequences, and more sensitive, allowing for genome-wide coverage with low input material and read numbers, in a fraction of the time.

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Mentioned in this Paper

Repetitive Region
Micrococcal Nuclease
Genome
Transcription, Genetic
Virus Replication
Genome Assembly Sequence
DNA-like RNA
Cleaved Cell
Hybrids
Genomics

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