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The DOI matches its Crossref record; 7 of the 7 references it cites were found in a registry or an index.

Direct observation of DNA target searching and cleavage by CRISPR-Cas12a

Jeon, Yongmoon, Choi, You Hee, Jang, Yunsu, Yu, Jihyeon, Goo, Jiyoung, Lee, Gyejun, Jeong, You Kyeong, Lee, Seung Hwan, Kim, In-San, Kim, Jin-Soo, Jeong, Cherlhyun, Lee, Sanghwa, Bae, Sangsu·2018·Nature Communications
DOI 10.1038/s41467-018-05245-xPMC PMC6050341
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Abstract

Cas12a (also called Cpf1) is a representative type V-A CRISPR effector RNA-guided DNA endonuclease, which provides an alternative to type II CRISPR–Cas9 for genome editing. Previous studies have revealed that Cas12a has unique features distinct from Cas9, but the detailed mechanisms of target searching and DNA cleavage by Cas12a are still unclear. Here, we directly observe this entire process by using single-molecule fluorescence assays to study Cas12a from Acidaminococcus sp. (AsCas12a). We determine that AsCas12a ribonucleoproteins search for their on-target site by a one-dimensional diffusion along elongated DNA molecules and induce cleavage in the two DNA strands in a well-defined order, beginning with the non-target strand. Furthermore, the protospacer-adjacent motif (PAM) for AsCas12a makes only a limited contribution of DNA unwinding during R-loop formation and shows a negligible role in the process of DNA cleavage, in contrast to the Cas9 PAM.

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Crossref · 10.1038/s41467-018-05245-x

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  • Cas12a differs from Cas9 by self-catalysis of crRNA maturation.

    “their distinguishable differences from Cas9 include: a T nucleotide-rich PAM site (5′-TTTV), self-catalysis of crRNA maturation, and a staggered dsDNA cut [76,77,78,79].”

    From Viral Delivery Systems for CRISPR (2019) · cites this paper as [79]

  • Cas12a differs from Cas9 by a T nucleotide-rich PAM site (5′-TTTV).

    “their distinguishable differences from Cas9 include: a T nucleotide-rich PAM site (5′-TTTV), self-catalysis of crRNA maturation, and a staggered dsDNA cut [76,77,78,79].”

    From Viral Delivery Systems for CRISPR (2019) · cites this paper as [79]

  • Cas12a differs from Cas9 by a staggered dsDNA cut.

    “their distinguishable differences from Cas9 include: a T nucleotide-rich PAM site (5′-TTTV), self-catalysis of crRNA maturation, and a staggered dsDNA cut [76,77,78,79].”

    From Viral Delivery Systems for CRISPR (2019) · cites this paper as [79]

  • Cas12a enzymes are single crRNA-guided endonucleases measuring 3.9 kb.

    “Cas12a enzymes are single crRNA-guided endonucleases measuring 3.9 kb”

    From Viral Delivery Systems for CRISPR (2019) · cites this paper as [79]

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Citations

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Metadata

Authors
Jeon, Y., Choi, Y. H., Jang, Y., Yu, J., Goo, J., Lee, G., Jeong, Y. K., Lee, S. H., Kim, I.-S., Kim, J.-S., Jeong, C., Lee, S., & Bae, S.
ORCID
Venue
Nature Communications
Publisher
Springer Science and Business Media LLC
Published
2018-07-17
Volume
9
Issue
1
ISSN
2041-1723
DOI
10.1038/s41467-018-05245-x
PMC
PMC6050341
Cited by
239
References
7
Language
en
Licence

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