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Updated: Feb 1, 2026

A Customizable Protocol for String Assembly gRNA Cloning STAgR
Published on: December 26, 2018
A novel cloning strategy for one-step assembly of multiplex CRISPR vectors
Marc Zuckermann1,2, Mario Hlevnjak3, Haniyeh Yazdanparast3
1Division of Molecular Genetics, German Cancer Research Center (DKFZ), Im Neuenheimer Feld 280, 69120, Heidelberg, Germany. M.Zuckermann@dkfz-heidelberg.de.
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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