ArticleMethods in molecular biology (Clifton, N.J.)2027
Genome-Wide Mapping of R-Loops in S. cerevisiae by H-CRAC.
Article in Methods in molecular biology (Clifton, N.J.), 2027. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Not yet cited in PubMed.
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Abstract
R-loops are noncanonical nucleic acid structures formed by an RNA:DNA heteroduplex, presumably arising by the pairing of the nascent RNA to its own DNA template and the subsequent extrusion of a single DNA strand. Such structures, previously thought to be rare and mostly by-products of transcription, are today recognized as physiological intermediates of several molecular processes and to form at much higher rates than anticipated. R-loop levels must be meticulously controlled, as they can lead to DNA damage and genome instability. RNA remodeling, in particular, constitutes a central mechanism for preventing and resolving R-loops. In this chapter, we describe H-CRAC, a procedure to chart R-loops genome-wide, in a strand-specific and high-resolution manner. H-CRAC relies on the capture and sequencing of RNAs UV-crosslinked to tagged RNases H, enzymes that selectively bind and digest RNA:DNA hybrids. In vivo genome-wide profiling of R-loops by H-CRAC reveals unprecedented sensitivity and resolution.
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