Open Access
Multiplex gene editing by CRISPR–Cpf1 using a single crRNA array
Bernd Zetsche,Matthias Heidenreich,Prarthana Mohanraju,Iana Fedorova,Jeroen Kneppers,Jeroen Kneppers,Ellen M DeGennaro,Ellen M DeGennaro,Nerges Winblad,Sourav R Choudhury,Omar O. Abudayyeh,Jonathan S. Gootenberg,Wen Y Wu,David A. Scott,David A. Scott,Konstantin Severinov,Konstantin Severinov,Konstantin Severinov,John van der Oost,Feng Zhang +19 more
TLDR
In the version of this article initially published, the percentage for the targets Mecp2, Nlgn3, and Drd1 should be 15.2%, not 16.9%; the same error appeared in the main text, next to last paragraph, “The authors' results show that ∼17%.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.read more
Citations
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Journal ArticleDOI
Efficient targeted DNA editing and replacement in Chlamydomonas reinhardtii using Cpf1 ribonucleoproteins and single-stranded DNA
TL;DR: It is reported that single-step codelivery of CRISPR/Cpf1 ribonucleoproteins with single-stranded DNA repair templates results in precise and targeted DNA replacement with as much as ∼10% efficiency in C. reinhardtii.
Journal ArticleDOI
dCas9-based epigenome editing suggests acquisition of histone methylation is not sufficient for target gene repression.
Henriette O'Geen,Chonghua Ren,Chonghua Ren,Charles M Nicolet,Andrew A. Perez,Julian Halmai,Victoria M. Le,Joel P. Mackay,Peggy J. Farnham,David J. Segal +9 more
TL;DR: The results suggest that so-called repressive histone modifications are not sufficient for gene repression, and the easily programmable dCas9 toolkit allowed precise control of epigenetic information and dissection of the relationship between the epigenome and gene regulation.
Journal ArticleDOI
Multiplex gene regulation by CRISPR-ddCpf1
Xiaochun Zhang,Xiaochun Zhang,Jingman Wang,Jingman Wang,Qiu-Xiang Cheng,Xuan Zheng,Guoping Zhao,Jin Wang +7 more
TL;DR: It is demonstrated that ddCpf1 alone could be employed for gene repression in Escherichia coli, and the repression was more effective with CRISPR RNAs (crRNAs) specifically targeting to the template strand of its target genes, which was different from that of dCas9.
Journal ArticleDOI
Immunity to CRISPR Cas9 and Cas12a therapeutics
TL;DR: Ways to assay and reduce the immunogenicity of Cas9 and Cas12a proteins are critical for ensuring patient safety and treatment efficacy, and for bringing us closer to realizing the vision of permanent genetic cures.
Journal ArticleDOI
CRISPR Cpf1 proteins: structure, function and implications for genome editing.
TL;DR: The evolutionary origins, basic architectures, and molecular mechanisms of Cpf1 family proteins, as well as crRNA designing and delivery strategies are discussed, which have broadened the versatility and feasibility of this system in genome editing, transcription regulation, epigenetic modulation, and base editing.
References
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Journal ArticleDOI
Multiplex gene editing by CRISPR-Cpf1 using a single crRNA array
Bernd Zetsche,Matthias Heidenreich,Prarthana Mohanraju,Iana Fedorova,Jeroen Kneppers,Jeroen Kneppers,Ellen M DeGennaro,Ellen M DeGennaro,Nerges Winblad,Sourav R Choudhury,Omar O. Abudayyeh,Jonathan S. Gootenberg,Wen Y Wu,David A. Scott,David A. Scott,Konstantin Severinov,Konstantin Severinov,Konstantin Severinov,John van der Oost,Feng Zhang +19 more
TL;DR: It is shown that the ability of Cpf1 to process its own CRISPR RNA (crRNA) can be used to simplify multiplexed genome editing.