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Scoop.it!
SITE-Seq probes Cas9 cleavage sites in vitro and returns a comprehensive list of off-target sites at different Cas9-sgRNA concentrations. 
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Scoop.it!
BigField GEG Tech's insight:
Often it can be difficult to identify a gene or genetic region that underlies a specific trait. Traditional mapping relies on naturally occurring recombination events but can be limited in resolution by the natural recombination frequency. Sadhu et al. used CRISPR technology to more easily map genomic sites of interest in yeast. The method systematically introduces local recombination events, allowing for the fine mapping of trait variants. They used their method to identify the causative mutation responsible for altered manganese sensitivity in yeast.
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Here, the authors developed a biochemical method (SITE-Seq), using Cas9 programmed with single-guide RNAs (sgRNAs), to identify the sequence of cut sites within genomic DNA. Cells edited with the same Cas9–sgRNA complexes are then assayed for mutations at each cut site using amplicon sequencing.