Evidence map›Paper›PMID 27342128›Full record

ArticleCell biochemistry and biophysics2017

Nucleic Acid-Dependent Conformational Changes in CRISPR-Cas9 Revealed by Site-Directed Spin Labeling.

Carolina Vazquez Reyes, Narin S Tangprasertchai, S D Yogesha, Richard H Nguyen, Xiaojun Zhang, Rakhi Rajan, Peter Z Qin

Abstract read
PubMed Full text at PMC Publisher
In one paragraph

Article in Cell biochemistry and biophysics, 2017. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 8 papers.

0numbers the graph read from it
0cells of the map it votes in
8citing papers in PubMed
–field-weighted citation impact
1 · What the graph read from it

What it found

Each row is one number read from the abstract, on the scale the paper reported it, with its interval. Left of the dashed line favours the treatment, right favours the comparator. Under each row is the sentence it came from. New to these charts? A ten-minute tutorial.

The abstract states no effect estimate the extractor could read, or names no intervention and outcome on the map, so this paper lights no cell and moves no belief. It is still indexed, cited and linked below.

2 · The registry

The trial behind it

Trials whose registry record cites this paper, or whose number appears in the abstract. A trial that started after this paper was published is citing it as background, not reporting it.

Neither the registry nor the abstract names a trial number. If this is a trial report, that itself is worth knowing.

3 · Its place in the literature

Who cites it

8 citing papers in PubMed.

  1. Review
  2. Article
  3. Article
  4. Article
  5. Review
  6. Article
  7. Article
  8. Article
4 · The record

Corrections and comments

PubMed lists nothing against this paper. Absence here is not a guarantee, only a check that was made.

5 · Who and what money

Authors and funding

7 authors.

Carolina Vazquez ReyesDepartment of Chemistry, University of Southern California, Los Angeles, CA, 90089, USA.
Narin S TangprasertchaiDepartment of Chemistry, University of Southern California, Los Angeles, CA, 90089, USA.
S D YogeshaDepartment of Chemistry and Biochemistry, University of Oklahoma, Norman, OK, 73019, USA.
Richard H NguyenDepartment of Chemistry and Biochemistry, University of Oklahoma, Norman, OK, 73019, USA.
Xiaojun ZhangDepartment of Chemistry, University of Southern California, Los Angeles, CA, 90089, USA.
Rakhi RajanDepartment of Chemistry and Biochemistry, University of Oklahoma, Norman, OK, 73019, USA.
Peter Z QinDepartment of Chemistry, University of Southern California, Los Angeles, CA, 90089, USA. pzq@usc.edu.

Funding

Structure-Function Studies of MsvR, a Methanogen-Specific Transcriptional RegulaP20GM103640 · NIGMS · UNIVERSITY OF OKLAHOMA · PI THOMAS, LEONARD M · 2012 to 2021
$20.4M
Acquisition of a Pulse Electron Paramagnetic Resonance SpectrometerS10RR028992 · NCRR · UNIVERSITY OF SOUTHERN CALIFORNIA · PI QIN, PETER Z · 2010 to 2010
$1.0M
NCRR NIH HHS S10 RR028992NIGMS NIH HHS P20 GM103640
6 · The paper itself

Abstract

In a type II clustered regularly interspaced short palindromic repeats (CRISPR) system, RNAs that are encoded at the CRISPR locus complex with the CRISPR-associated (Cas) protein Cas9 to form an RNA-guided nuclease that cleaves double-stranded DNAs at specific sites. In recent years, the CRISPR-Cas9 system has been successfully adapted for genome engineering in a wide range of organisms. Studies have indicated that a series of conformational changes in Cas9, coordinated by the RNA and the target DNA, direct the protein into its active conformation, yet details on these conformational changes, as well as their roles in the mechanism of function of Cas9, remain to be elucidated. Here, nucleic acid-dependent conformational changes in Streptococcus pyogenes Cas9 (SpyCas9) were investigated using the method of site-directed spin labeling (SDSL). Single nitroxide spin labels were attached, one at a time, at one of the two native cysteine residues (Cys80 and Cys574) of SpyCas9, and the spin-labeled proteins were shown to maintain their function. X-band continuous-wave electron paramagnetic resonance spectra of the nitroxide attached at Cys80 revealed conformational changes of SpyCas9 that are consistent with a large-scale domain re-arrangement upon binding to its RNA partner. The results demonstrate the use of SDSL to monitor conformational changes in CRISPR-Cas9, which will provide key information for understanding the mechanism of CRISPR function.

Indexed as

Clustered Regularly Interspaced Short Palindromic RepeatsCRISPR-Cas SystemsBacterial ProteinsCloning, MolecularCRISPR-Associated Protein 9DNAElectron Spin Resonance SpectroscopyEndonucleasesEscherichia coliGene ExpressionModels, MolecularNitrogen OxidesProtein BindingProtein DomainsProtein Structure, SecondaryRecombinant ProteinsBacterial ProteinsCas9 endonuclease Streptococcus pyogenesCRISPR-Associated Protein 9DNAEndonucleasesNitrogen OxidesnitroxylRecombinant ProteinsRNA, Guide, CRISPR-Cas SystemsSpin LabelsCas9Conformational changeCRISPREPRSpin labeling

Identifiers

PMID27342128
PMCPMC5183522

What OpenQuestion holds

Textmetadata
LicenceTDM
Read underepoch 390

Registered trials

None linked

Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.