High-Throughput mapping of the regulatory DNA

ABSTRACT: We introduce the Multiplexed Editing Regulatory Assay (MERA), a high-throughput CRISPR/Cas9-based approach that analyzes the regulatory genome for function in its native context. By tiling thousands of mutations across ~40 kb of cis-regulatory genomic space and using knock-in GFP reporters to read out gene activity, we obtain quantitative information on the contribution of cis-regulatory regions to gene expression. In addition, we performed a deep-sequencing strategy to find basepair-resolution functional motifs involved in regulation of the gene by sequencing thousands of functional and non-functional genotypes at genomic locations perturbed by specific guide RNAs. Design of 3908 gRNAs to perturb regulatory regions associated with the genes Tdgf1,Nanog,Zfp42 and Rpp25 as well as 10 GFP

ORGANISM(S): Mus musculus

SUBMITTER: Nisha Rajagopal 

PROVIDER: E-GEOD-76318 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress