Journal Article

DKFZ-2019-00016

http://join2-wiki.gsi.de/foswiki/pub/Main/Artwork/join2_logo100x88.png A novel cloning strategy for one-step assembly of multiplex CRISPR vectors.

Zuckermann, M. (First author)DKFZ* ; Hlevnjak, M.DKFZ* ; Yazdanparast, H.DKFZ* ; Zapatka, M.DKFZ* ; Jones, D.DKFZ* ; Lichter, P.DKFZ* ; Gronych, J. (Last author)DKFZ*

2018
Macmillan Publishers Limited, part of Springer Nature [London]

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Please use a persistent id in citations: doi:10.1038/s41598-018-35727-3

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.

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Contributing Institute(s):

1. B060 Molekulare Genetik (B060)
2. B062 Pädiatrische Neuroonkologie (B062)
3. DKTK Heidelberg (L101)

Research Program(s):

1. 312 - Functional and structural genomics (POF3-312) (POF3-312)

Appears in the scientific report 2018

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