Genome-wide specificities of CRISPR-Cas Cpf1 nucleases in human cells
By B. Kleinstiver, S. Tsai, Michelle S. Prew, Nhu T. Nguyen, Moira M. Welch, J. M. López, Z. McCaw, Martin J. Aryee, J. Keith Joung. Nature Biotechnology, 2016
645 citations recorded by the source corpus.
The activities and genome-wide specificities of CRISPR-Cas Cpf1 nucleases are not well defined. We show that two Cpf1 nucleases from Acidaminococcus sp. BV3L6 and Lachnospiraceae bacterium ND2006 (AsCpf1 and LbCpf1, respectively) have on-target efficiencies in human cells comparable with those of the widely used Streptococcus pyogenes Cas9 (SpCas9). We also report that four to six bases at the 3′ end of the short CRISPR RNA (crRNA) used to program Cpf1 nucleases are insensitive to single base mismatches, but that many of the other bases in this region of the crRNA are highly sensitive to single or double substitutions. Using GUIDE-seq and targeted deep sequencing analyses performed with both Cpf1 nucleases, we were unable to detect off-target cleavage for more than half of 20 different crRNAs. Our results suggest that AsCpf1 and LbCpf1 are highly specific in human cells.
Subfields: Molecular Biology
View Paper: https://www.semanticscholar.org/paper/621f533181d3eaa126ef77bbcba76e637fe208a1
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Citations in the corpus
- The Biology of CRISPR-Cas: Backward and Forward. (2018, neutral): “Moreover, Cas12a and Cas12b require a seed sequence of approximately 18 nt, implying that they could be highly specific alternatives to Cas9 for genome editing applications (Kim et al., 2016a, 2017; Kleinstiver et al., 2016; Liu et al., 2017a).”
- Diverse evolutionary roots and mechanistic variations of the CRISPR-Cas systems (2016, background): “Besides, Cpf1 is also highly specific in its targeting, as minimal or no off-target cleavage has been detected (153, 154).”
- Genome Editing—Principles and Applications for Functional Genomics Research and Crop Improvement (2017, background): “Studies in mammalian cells have shown that compared with CRISPR/Cas9, CRISPR-Cpf1 exhibited minimal off-target effects and from the limited published literature, it seems that is also the case in plants (Hur et al., 2016; Kim et al., 2016; Kleinstiver et al., 2016).”