Repair_profiles_in_DNA_repair_mutant_mouse_ES_cells_-sc-6243
Repair_profiles_in_DNA_repair_mutant_mouse_ES_cells_
We have developed a CRISPR/Cas9 platform for the analysis of DNA repair outcomes in mouse ES cells. Mouse embryonic stem cells that express Cas9 and have a mutation in a DNA repair pathway gene are infected with lentivirus that encodes for both a guide RNA and a target DNA sequence for that guide. After screening the cells for 2 weeks the genomic DNA is extracted and the target DNA is amplified by PCR and sequenced.   This data is part of a pre-publication release. For information on the proper use of pre-publication data shared by the Wellcome Trust Sanger Institute (including details of any publication moratoria), please see http://www.sanger.ac.uk/datasharing/
We have developed a CRISPR/Cas9 platform for the analysis of DNA repair outcomes in mouse ES cells. Sixteen Cas9-expressing mouse embryonic stem cell lines, each harboring a different mutation in a DNA repair pathway gene, were infected with lentivirus that encodes for both a guide RNA and a target DNA sequence for that guide. Two weeks later genomic DNA is extracted and the target DNA is amplified by PCR and sequenced. Deep sequencing data will reveal DNA repair profiles specific to each mutant line. The purpose of this study is to sequence the editing outcomes of 16 screens in duplicate at days 7 and 10 after infection.