The DOI matches its Crossref record; 18 of the 18 references it cites were found in a registry or an index.
Structural basis of PAM-dependent target DNA recognition by the Cas9 endonuclease
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Once the target is identified, the dsDNA is unwound and crRNA binds to the target ssDNA, leading to an R-loop shape and ultimately to a blunt double-strand break by both catalytic sites of Cas9, RuvC and HNH, 3 nt upstream of the PAM site.
“Once identified, the dsDNA is unwinded and crRNA binds to the target ssDNA leading to an R-loop shape and ultimately to a blunt double-strand break by both catalytic sites of Cas9, RuvC and HNH, 3 nt upstream of the PAM site [25,53,54].”
From CRISPR-Cas: Converting A Bacterial Defence Mechanism into A State-of-the-Art Genetic Manipulation Tool (2019) · cites this paper as [25]
The paper’s model (section labeled Abstract; page unavailable) describes PAM-dependent DNA unwinding and guide RNA–target DNA pairing, and it identifies HNH and RuvC as Cas9’s nuclease domains. Our copy does not state that their cleavage produces a blunt double-strand break three nucleotides upstream of the PAM; the structure described has an inactivated HNH domain and an intact target strand. (LLM, conf=0.84)
Experimental — accuracy not yet measuredgRNA molecules contain both a scaffold sequence that binds to Cas9 and a targeting sequence which directs the system towards the target locus.
“gRNA molecules contain both a scaffold sequence that binds to Cas9 and a targeting sequence which directs the system towards the target locus [25].”
From CRISPR-Cas: Converting A Bacterial Defence Mechanism into A State-of-the-Art Genetic Manipulation Tool (2019) · cites this paper as [25]
The paper’s Abstract (page unavailable) says Cas9 associates with guide RNA and that a 20-nucleotide segment of the guide RNA is complementary to target DNA, supporting the targeting-sequence part of the claim. The passages in our copy do not identify a scaffold sequence or state that it binds Cas9. (LLM, conf=0.82)
Experimental — accuracy not yet measuredIn type II systems, Cas9 recognizes the PAM sequence in the double-strand form.
“As for type II systems, Cas9 recognizes the PAM sequence in the double-strand form [25].”
From CRISPR-Cas: Converting A Bacterial Defence Mechanism into A State-of-the-Art Genetic Manipulation Tool (2019) · cites this paper as [25]
In type I systems, Cas1 binds to the PAM-complementary sequence in its ssDNA form.
“In type I systems, Cas1 binds to the PAM-complementary sequence in its ssDNA form.”
From CRISPR-Cas: Converting A Bacterial Defence Mechanism into A State-of-the-Art Genetic Manipulation Tool (2019) · cites this paper as [25]
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Metadata
- Authors
- Anders, C., Niewoehner, O., Duerst, A., & Jinek, M.
- Venue
- Nature
- Published
- 2014-07-27
- Volume
- 513
- Issue
- 7519
- Pages
- 569-573
- ISSN
- 0028-0836
- 1476-4687
- PMC
- PMC4176945
- Cited by
- 1,323
- References
- 18
- Language
- en
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- www.springer.com/tdm (tdm)
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