Student or educator? Sign up with your school email and get 6 months of Pro free.
CoChat
Sign Up
Cited by 365

The DOI matches its Crossref record; 13 of the 13 references it cites were found in a registry or an index.

Structure and Engineering of Francisella novicida Cas9

Open access
Hirano, Hisato, Gootenberg, Jonathan S., Horii, Takuro, Abudayyeh, Omar O., Kimura, Mika, Hsu, Patrick D., Nakane, Takanori, Ishitani, Ryuichiro, Hatada, Izuho, Zhang, Feng, Nishimasu, Hiroshi, Nureki, Osamu·2016·Cell
DOI 10.1016/j.cell.2016.01.039
Open this paper in CoChat — free account

Abstract

Summary The RNA-guided endonuclease Cas9 cleaves double-stranded DNA targets complementary to the guide RNA, and has been applied to programmable genome editing. Cas9-mediated cleavage requires a protospacer adjacent motif (PAM) juxtaposed with the DNA target sequence, thus constricting the range of targetable sites. Here, we report the 1.7 Å resolution crystal structures of Cas9 from Francisella novicida (FnCas9), one of the largest Cas9 orthologs, in complex with a guide RNA and its PAM-containing DNA targets. A structural comparison of FnCas9 with other Cas9 orthologs revealed striking conserved and divergent features among distantly related CRISPR-Cas9 systems. We found that FnCas9 recognizes the 5′-NGG-3′ PAM, and used the structural information to create a variant that can recognize the more relaxed 5′-YG-3′ PAM. Furthermore, we demonstrated that pre-assembled FnCas9 ribonucleoprotein complexes can be microinjected into mouse zygotes to edit endogenous sites with the 5′-YG-3′ PAMs, thus expanding the target space of the CRISPR-Cas9 toolbox.

Overview

The claims and statistics of this paper haven’t been checked yet.

Run the check — free, no account needed. It reads the open-access text and checks the sources it cites.

A few free checks a day · papers already verified open instantly · results are public

Integrity signals

Registry

Crossref · 10.1016/j.cell.2016.01.039

Cites

13 of 13 references verified0 citation errors · 0 not checkable

Statistics

Verification

Claims the paper makes

The claims and statistics of this paper haven’t been checked yet.

Run the check — free, no account needed. It reads the open-access text and checks the sources it cites.

A few free checks a day · papers already verified open instantly · results are public

Self-check

Percentages, sums and reported differences re-computed from the paper’s own numbers.

The self-check needs the paper’s full text.

Citations

Cites

Show all 13 references

Research works printed but not linked: 23

  1. Anders, 2014, Structural basis of PAM-dependent target DNA recognition by the Cas9 endonuclease, Nature, 513, 569, 10.1038/nature13579 · DOI · Paper page
  2. Briner, 2014, Guide RNA functional modules direct Cas9 activity and orthogonality, Mol. Cell, 56, 333, 10.1016/j.molcel.2014.09.019 · DOI · Paper page
  3. Chylinski, 2013, The tracrRNA and Cas9 families of type II CRISPR-Cas immunity systems, RNA Biol., 10, 726, 10.4161/rna.24321 · DOI · Paper page
  4. Cowtan, 2006, The Buccaneer software for automated model building. 1. Tracing protein chains, Acta Crystallogr. D Biol. Crystallogr., 62, 1002, 10.1107/S0907444906022116 · DOI · Paper page
  5. Crooks, 2004, WebLogo: a sequence logo generator, Genome Res., 14, 1188, 10.1101/gr.849004 · DOI · Paper page
  6. Deveau, 2008, Phage response to CRISPR-encoded resistance in Streptococcus thermophilus, J. Bacteriol., 190, 1390, 10.1128/JB.01412-07 · DOI · Paper page
  7. Evans, 2013, How good are my data and what is the resolution?, Acta Crystallogr. D Biol. Crystallogr., 69, 1204, 10.1107/S0907444913000061 · DOI · Paper page
  8. Fonfara, 2014, Phylogeny of Cas9 determines functional exchangeability of dual-RNA and Cas9 among orthologous type II CRISPR-Cas systems, Nucleic Acids Res., 42, 2577, 10.1093/nar/gkt1074 · DOI · Paper page
  9. Garneau, 2010, The CRISPR/Cas bacterial immune system cleaves bacteriophage and plasmid DNA, Nature, 468, 67, 10.1038/nature09523 · DOI · Paper page
  10. Gasiunas, 2012, Cas9-crRNA ribonucleoprotein complex mediates specific DNA cleavage for adaptive immunity in bacteria, Proc. Natl. Acad. Sci. USA, 109, E2579, 10.1073/pnas.1208507109 · DOI · Paper page
  11. Hsu, 2014, Development and applications of CRISPR-Cas9 for genome engineering, Cell, 157, 1262, 10.1016/j.cell.2014.05.010 · DOI · Paper page
  12. Jiang, 2015, A Cas9-guide RNA complex preorganized for target DNA recognition, Science, 348, 1477, 10.1126/science.aab1452 · DOI · Paper page
  13. Jiang, 2016, Structures of a CRISPR-Cas9 R-loop complex primed for DNA cleavage, Science, 10.1126/science.aad8282 · DOI · Paper page
  14. Kleinstiver, 2015, Broadening the targeting range of Staphylococcus aureus CRISPR-Cas9 by modifying PAM recognition, Nat. Biotechnol., 33, 1293, 10.1038/nbt.3404 · DOI · Paper page
  15. Lin, 2014, Enhanced homology-directed human genome engineering by controlled timing of CRISPR/Cas9 delivery, eLife, 3, e04766, 10.7554/eLife.04766 · DOI · Paper page
  16. Mojica, 2009, Short motif sequences determine the targets of the prokaryotic CRISPR defence system, Microbiology, 155, 733, 10.1099/mic.0.023960-0 · DOI · Paper page
  17. Nishimasu, 2015, Crystal structure of Staphylococcus aureus Cas9, Cell, 162, 1113, 10.1016/j.cell.2015.08.007 · DOI · Paper page
  18. Sampson, 2013, A CRISPR/Cas system mediates bacterial innate immune evasion and virulence, Nature, 497, 254, 10.1038/nature12048 · DOI · Paper page
  19. Sternberg, 2015, Conformational control of DNA target cleavage by CRISPR-Cas9, Nature, 527, 110, 10.1038/nature15544 · DOI · Paper page
  20. Sung, 2014, Highly efficient gene knockout in mice and zebrafish with RNA-guided endonucleases, Genome Res., 24, 125, 10.1101/gr.163394.113 · DOI · Paper page
  21. Woo, 2015, DNA-free genome editing in plants with preassembled CRISPR-Cas9 ribonucleoproteins, Nat. Biotechnol., 33, 1162, 10.1038/nbt.3389 · DOI · Paper page
  22. Zetsche, 2015, Cpf1 is a single RNA-guided endonuclease of a class 2 CRISPR-Cas system, Cell, 163, 759, 10.1016/j.cell.2015.09.038 · DOI · Paper page
  23. Zuris, 2015, Cationic lipid-mediated delivery of proteins enables efficient protein-based genome editing in vitro and in vivo, Nat. Biotechnol., 33, 73, 10.1038/nbt.3081 · DOI · Paper page

Cited by

Metadata

Authors
Hirano, H., Gootenberg, J. S., Horii, T., Abudayyeh, O. O., Kimura, M., Hsu, P. D., Nakane, T., Ishitani, R., Hatada, I., Zhang, F., Nishimasu, H., & Nureki, O.
Venue
Cell
Publisher
Elsevier BV
Published
2016-02
Volume
164
Issue
5
Pages
950-961
ISSN
0092-8674 (ISSN-L)
DOI
10.1016/j.cell.2016.01.039
OpenAlex
W2268491398
Cited by
365
References
13
FWCI
21.67
Type
journal article
Language
en
Open access
bronze
Licence
Funders
  • D.O.E. Computational Science Graduate Fellowship
  • Japan Agency for Medical Research and Development
  • NIH through NIMH (5DP1-MH100706, 1R01-MH110049)
  • NIDDK (5R01DK097768-03)
  • New York Stem Cell
  • Simons
  • Paul G. Allen Family
  • Vallee Foundations
  • JST, PRESTO
  • JSPS KAKENHI (26291010, 15H01463)
  • Ministry of Education, Culture, Sports, Science, and Technology

About these checks

Registry, retraction, Cites and statistics results are computed, not judged, with one exception: a printed reference that no index lists is read once by a model before it counts as a citation error. Claims are extracted by a model and kept only when their quote appears verbatim in the text; each cited source is then checked, with a model’s help, for whether it says what the claim relies on. Those verdicts can be wrong — read the claim’s quote and the cited source before relying on one.

Statistics and arithmetic are recomputed, not judged.

Confidence bands are shown only for fields calibrated on our annotated test set. None is calibrated yet, so no band is shown.

What counts as a claim ›How we measure confidence ›How we check this ›

Are you an author, or do you see a problem with this page? Request a review

Data sources

Bibliographic data: Semantic Scholar, OpenAlex, Crossref.

Contains information from the Semantic Scholar Academic Graph Datasets, made available under the ODC Attribution License (ODC-BY 1.0).

Semantic Scholar’s data is described in: Kinney, R., et al. (2023). The Semantic Scholar Open Data Platform. arXiv:2301.10140.