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Multiplex gene editing by CRISPR–Cpf1 using a single crRNA array

2016; Nature Portfolio; Volume: 35; Issue: 1 Linguagem: Inglês

10.1038/nbt.3737

1546-1696

Bernd Zetsche, Matthias Heidenreich, Prarthana Mohanraju, Iana Fedorova, Jeroen Kneppers, Ellen M. DeGennaro, Nerges Winblad, Sourav Choudhury, Omar O. Abudayyeh, Jonathan S. Gootenberg, Wen Y. Wu, David Scott, Konstantin Severinov, John van der Oost, Feng Zhang,

RNA and protein synthesis mechanisms

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.