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DNA interrogation by the CRISPR RNA-guided endonuclease Cas9

Overview of attention for article published in Nature, January 2014
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  • In the top 5% of all research outputs scored by Altmetric
  • High Attention Score compared to outputs of the same age (99th percentile)
  • High Attention Score compared to outputs of the same age and source (82nd percentile)

Citations

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1551 Dimensions

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3460 Mendeley
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7 CiteULike
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Title
DNA interrogation by the CRISPR RNA-guided endonuclease Cas9
Published in
Nature, January 2014
DOI 10.1038/nature13011
Pubmed ID
Authors

Samuel H. Sternberg, Sy Redding, Martin Jinek, Eric C. Greene, Jennifer A. Doudna

Abstract

The clustered regularly interspaced short palindromic repeats (CRISPR)-associated enzyme Cas9 is an RNA-guided endonuclease that uses RNA-DNA base-pairing to target foreign DNA in bacteria. Cas9-guide RNA complexes are also effective genome engineering agents in animals and plants. Here we use single-molecule and bulk biochemical experiments to determine how Cas9-RNA interrogates DNA to find specific cleavage sites. We show that both binding and cleavage of DNA by Cas9-RNA require recognition of a short trinucleotide protospacer adjacent motif (PAM). Non-target DNA binding affinity scales with PAM density, and sequences fully complementary to the guide RNA but lacking a nearby PAM are ignored by Cas9-RNA. Competition assays provide evidence that DNA strand separation and RNA-DNA heteroduplex formation initiate at the PAM and proceed directionally towards the distal end of the target sequence. Furthermore, PAM interactions trigger Cas9 catalytic activity. These results reveal how Cas9 uses PAM recognition to quickly identify potential target sites while scanning large DNA molecules, and to regulate scission of double-stranded DNA.

X Demographics

X Demographics

The data shown below were collected from the profiles of 75 X users who shared this research output. Click here to find out more about how the information was compiled.
Mendeley readers

Mendeley readers

The data shown below were compiled from readership statistics for 3,460 Mendeley readers of this research output. Click here to see the associated Mendeley record.

Geographical breakdown

Country Count As %
United States 45 1%
United Kingdom 13 <1%
Germany 12 <1%
Spain 6 <1%
China 5 <1%
Denmark 4 <1%
Japan 3 <1%
Netherlands 3 <1%
Mexico 2 <1%
Other 21 <1%
Unknown 3346 97%

Demographic breakdown

Readers by professional status Count As %
Student > Ph. D. Student 815 24%
Student > Bachelor 566 16%
Researcher 517 15%
Student > Master 477 14%
Student > Doctoral Student 143 4%
Other 401 12%
Unknown 541 16%
Readers by discipline Count As %
Agricultural and Biological Sciences 1212 35%
Biochemistry, Genetics and Molecular Biology 1083 31%
Medicine and Dentistry 115 3%
Chemistry 96 3%
Engineering 81 2%
Other 287 8%
Unknown 586 17%
Attention Score in Context

Attention Score in Context

This research output has an Altmetric Attention Score of 181. This is our high-level measure of the quality and quantity of online attention that it has received. This Attention Score, as well as the ranking and number of research outputs shown below, was calculated when the research output was last mentioned on 05 March 2024.
All research outputs
#221,566
of 25,377,790 outputs
Outputs from Nature
#12,937
of 97,788 outputs
Outputs of similar age
#2,008
of 323,417 outputs
Outputs of similar age from Nature
#160
of 888 outputs
Altmetric has tracked 25,377,790 research outputs across all sources so far. Compared to these this one has done particularly well and is in the 99th percentile: it's in the top 5% of all research outputs ever tracked by Altmetric.
So far Altmetric has tracked 97,788 research outputs from this source. They typically receive a lot more attention than average, with a mean Attention Score of 102.4. This one has done well, scoring higher than 86% of its peers.
Older research outputs will score higher simply because they've had more time to accumulate mentions. To account for age we can compare this Altmetric Attention Score to the 323,417 tracked outputs that were published within six weeks on either side of this one in any source. This one has done particularly well, scoring higher than 99% of its contemporaries.
We're also able to compare this research output to 888 others from the same source and published within six weeks on either side of this one. This one has done well, scoring higher than 82% of its contemporaries.