A novel cloning strategy for one-step assembly of multiplex CRISPR vectors. Issue 1 (December 2018)

• Record Type: Journal Article
• Authors: Zuckermann, Marc; Hlevnjak, Mario; Yazdanparast, Haniyeh; Zapatka, Marc; Jones, David; Lichter, Peter; Gronych, Jan
• Is Part Of: Scientific reports. Volume 8:Issue 1(2018)
• Page Start: 1
• Page End: 8
• Publication Date: 2018-12
• Subjects: Natural history -- Research -- Periodicals; Biology -- Research -- Periodicals; Physical sciences -- Research -- Periodicals; Earth sciences -- Research -- Periodicals; Environmental sciences -- Research -- Periodicals; 502.85
• DOI: 10.1038/s41598-018-35727-3
• Languages: English
• ISSNs: 2045-2322

Abstract

Abstract One key advantage of the CRISPR/Cas9 system in comparison with other gene editing approaches lies in its potential for multiplexing. Here, we describe an elaborate procedure that allows the assembly of multiple gRNA expression cassettes into a vector of choice within a single step, termed ASAP(Adaptable System for Assembly of multiplexed Plasmids)-cloning. We demonstrate the utility of ASAP-cloning for multiple CRISPR-mediated applications, including efficient multiplex gene editing, robust transcription activation and convenient analysis of Cas9 activity in the presence of multiple gRNAs.