title: Selection and Validation of Spacer Sequences for CRISPR-Cas9 Genome Editing and Transcription Regulation in Bacteria.
abstract: RNA-guided Cas9 nucleases derived from clustered regularly interspaced short palindromic repeat (CRISPR)-Cas systems have recently been adapted as sequence-programmable tools for various purposes such as genome editing and transcriptional regulation. A critical aspect of the system is the selection and validation of spacer sequences that allow precise targeting of the guide RNA-Cas9 complex. We describe a procedure involving computational and experimental steps to identify and test potentially interesting spacer sequences in bacterial genomes.
authors.affiliation: Département de Biologie, Faculté des Sciences, Université de Sherbrooke, 2500 boulevard de l'Université, Sherbrooke, QC, Canada, J1K 2R1.
authors.forename: Frédéric
authors.initials: F
authors.lastname: Grenier
authors.affiliation: Département de Biologie, Faculté des Sciences, Université de Sherbrooke, 2500 boulevard de l'Université, Sherbrooke, QC, Canada, J1K 2R1.
authors.forename: Jean-François
authors.initials: JF
authors.lastname: Lucier
authors.affiliation: Département de Biologie, Faculté des Sciences, Université de Sherbrooke, 2500 boulevard de l'Université, Sherbrooke, QC, Canada, J1K 2R1. sebastien.rodrigue@usherbrooke.ca.
authors.forename: Sébastien
authors.initials: S
authors.lastname: Rodrigue
mesh_terms: D064113:CRISPR-Cas Systems / Q000235:genetics; D064112:Clustered Regularly Interspaced Short Palindromic Repeats / Q000235:genetics*; D004720:Endonucleases / Q000235:genetics; D005786:Gene Expression Regulation; D005818:Genetic Engineering / Q000379:methods*; D016680:Genome, Bacterial; D009154:Mutation; D017393:RNA Editing / Q000235:genetics*; D014158:Transcription, Genetic
chemical_list: D004720:Endonucleases
keywords: CRISPR; Cas9; Genome editing; Repression; Transcription; gRNA