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CRISPR/Cas9: Comparison
Please note this is a comparison between Version 3 by Yu Peng and Version 2 by Catherine Yang.

CRISPR/Cas9 is an RNA-guided DNA-targeting system derived from the type II CRISPR adaptive immune systems of bacteria. Its core components are the Cas9 endonuclease and a guide RNA, which may consist of separate CRISPR RNA (crRNA) and trans-activating crRNA (tracrRNA) molecules or a fused single-guide RNA (sgRNA) [1]. Base pairing between the guide-derived spacer sequence and a complementary DNA protospacer directs Cas9 to a specified genomic or extrachromosomal locus. Target recognition additionally requires a nearby protospacer-adjacent motif (PAM), whose sequence is determined by the particular Cas9 ortholog; for the commonly used Streptococcus pyogenes Cas9, this motif is generally 5′-NGG-3′ [1][2]. After PAM recognition and guide–target hybridization, the HNH and RuvC nuclease domains of Cas9 cleave opposite DNA strands to generate a double-strand break [1][3]. CRISPR/Cas9 therefore denotes the guide-RNA-programmable Cas9 system itself, rather than the broader CRISPR family, which includes mechanistically distinct class 1 and class 2 CRISPR systems and non-Cas9 effector proteins [3].

  • Cas9 endonuclease
  • single-guide RNA
  • protospacer-adjacent motif
  • RNA-guided genome editing

References

  1. Martin Jinek; Krzysztof Chylinski; Ines Fonfara; Michael Hauer; Jennifer A. Doudna; Emmanuelle Charpentier; A Programmable Dual-RNA–Guided DNA Endonuclease in Adaptive Bacterial Immunity. Science 2012, 337, 816-821. [CrossRef]
  2. Giedrius Gasiunas; Rodolphe Barrangou; Philippe Horvath; Virginijus Siksnys; Cas9–crRNA ribonucleoprotein complex mediates specific DNA cleavage for adaptive immunity in bacteria. Proc. Natl. Acad. Sci. 2012, 109, E2579-E2586. [CrossRef]
  3. Frank Hille; Hagen Richter; Shi Pey Wong; Majda Bratovič; Sarah Ressel; Emmanuelle Charpentier; The Biology of CRISPR-Cas: Backward and Forward. Cell 2018, 172, 1239-1259. [CrossRef]
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