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Nat Biotechnol. 2017 Jan;35(1):31-34. doi: 10.1038/nbt.3737. Epub 2016 Dec 5.

Multiplex gene editing by CRISPR-Cpf1 using a single crRNA array.

Zetsche B#1,2,3,4,5, Heidenreich M#1,2,3,4, Mohanraju P#6, Fedorova I1,2,3,4,7,8, Kneppers J1,6, DeGennaro EM1,9, Winblad N1,2,3,4, Choudhury SR1,2,3,4, Abudayyeh OO1,2,3,4,9, Gootenberg JS1,2,3,4,10, Wu WY6, Scott DA1,2, Severinov K7,11,12, van der Oost J6, Zhang F1,2,3,4.

Author information

1
Broad Institute of MIT and Harvard, Cambridge, MA 02142.
2
McGovern Institute for Brain Research, Massachusetts Institute of Technology, Cambridge, MA 02139.
3
Department of Brain and Cognitive Sciences, Massachusetts Institute of Technology, Cambridge, MA 02139.
4
Department of Biological Engineering, Massachusetts Institute of Technology, Cambridge, MA 02139.
5
Department of Developmental Pathology, Institute of Pathology, Bonn Medical School, Sigmund Freud Street 25, 53127 Bonn, Germany.
6
Laboratory of Microbiology, Department of Agrotechnology and Food Sciences, Wageningen University, Stippeneng 4, 6708 WE Wageningen, Netherlands.
7
Skolkovo Institute of Science and Technology, Skolkovo, 143025, Russia.
8
Peter the Great St.Petersburg Polytechinc University, St. Petersburg, 195251, Russia.
9
Harvard-MIT Division of Health Sciences and Technology, Massachusetts Institute of Technology, Cambridge, MA 02139.
10
Department of Systems Biology, Harvard Medical School, Boston, MA 02115.
11
Waksman Institute for Microbiology, Rutgers, The State University of New Jersey, Piscataway, NJ 08854, USA.
12
Institute of Molecular Genetics, Russian Academy of Sciences, Moscow, 123182, Russia.
#
Contributed equally

Abstract

Targeting of multiple genomic loci with Cas9 is limited by the need for multiple or large expression constructs. Here we show that the ability of Cpf1 to process its own CRISPR RNA (crRNA) can be used to simplify multiplexed genome editing. Using a single customized CRISPR array, we edit up to four genes in mammalian cells and three in the mouse brain, simultaneously.

PMID:
27918548
PMCID:
PMC5225075
DOI:
10.1038/nbt.3737
[Indexed for MEDLINE]
Free PMC Article

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