title: Efficient programmable gene silencing by Cascade.
abstract: Methods that permit controlled changes in the expression of genes are important tools for biological and medical research, and for biotechnological applications. Conventional methods are directed at individually changing each gene, its regulatory elements or its mRNA's translation rate. We demonstrate that the CRISPR-associated DNA-binding Cascade complex can be used for efficient, long-lasting and programmable gene silencing. When Cascade is targeted to a promoter sequence the transcription of the downstream gene is inhibited, resulting in dramatically reduced expression. The specificity of Cascade binding is provided by the integral crRNA component, which is easily designed to target virtually any stretch of DNA. Cascade targeted to the ORF sequence of the gene can also silence expression, albeit at lower efficiency. The system can be used to silence plasmid and chromosome targets, simultaneously target several genes and is active in different bacterial species and strains. The findings described here are an addition to the expanding range of CRISPR-based technologies and may be adapted to additional organisms and cell systems.
authors.affiliation: Department of Cell and Molecular Biology, Uppsala University, SE-751 24 Uppsala, Sweden.
authors.forename: Devashish
authors.initials: D
authors.lastname: Rath
authors.affiliation: Department of Cell and Molecular Biology, Uppsala University, SE-751 24 Uppsala, Sweden.
authors.forename: Lina
authors.initials: L
authors.lastname: Amlinger
authors.affiliation: Department of Cell and Molecular Biology, Uppsala University, SE-751 24 Uppsala, Sweden.
authors.forename: Mirthe
authors.initials: M
authors.lastname: Hoekzema
authors.affiliation: Department of Cell and Molecular Biology, Uppsala University, SE-751 24 Uppsala, Sweden.
authors.forename: Praneeth Reddy
authors.initials: PR
authors.lastname: Devulapally
authors.affiliation: Department of Cell and Molecular Biology, Uppsala University, SE-751 24 Uppsala, Sweden magnus.lundgren@icm.uu.se.
authors.forename: Magnus
authors.initials: M
authors.lastname: Lundgren
mesh_terms: D064130:CRISPR-Associated Proteins / Q000378:metabolism*; D004269:DNA, Bacterial / Q000378:metabolism; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D020868:Gene Silencing; D008957:Models, Genetic; D016366:Open Reading Frames; D011401:Promoter Regions, Genetic; D012329:RNA, Bacterial / Q000378:metabolism*; D014158:Transcription, Genetic
chemical_list: D064130:CRISPR-Associated Proteins; D004269:DNA, Bacterial; D012329:RNA, Bacterial