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==CRISPR Cas9 interaction with target DNA== | ==CRISPR Cas9 interaction with target DNA== | ||
he CRISPR/Cas9 complex of Streptococcus pyogenes consists of the Cas9 protein (in gray) and a guide RNA that is a chimera of natural crRNA and tracRNA (in orange). | |||
The targeting sequence at the 5′end of the guide RNA base-pairs with complementary sequences on the target DNA (in blue); the targeting sequence is 20 nucleotides long, but may be shortened to increase specificity (the addition of 1 to 2 unpaired nucleotides at the 5′ end is also tolerated). | |||
The presence of a PAM (protospacer adjacent motif, NGG for Streptococcus pyogenes), located immediately downstream of the 20-nucleotide sequence targeted by the guide RNA, is also essential for target recognition and cleavage. | |||
The PAM sequence does not have a counterpart on the guide RNA. Following recognition of the PAM and base-pairing between the guide RNA and the target, | |||
Cas9 cleaves each of the target DNA strands a few nucleotides upstream of the PAM (red arrowheads). Each strand is cleaved by a different nuclease domain present in Cas9 (HNH and RuvC domains). These domains have been mutated independently to generate Cas9 nickases. | |||
===Reference=== | |||
<pubmed>25699168</pubmed>| [http://www.evodevojournal.com/content/5/1/43 Evodevo.] | |||
====Copright==== | |||
© 2014 Gilles and Averof; licensee BioMed Central Ltd. | |||
This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly credited. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated. | |||
Gilles and Averof EvoDevo 2014 5:43 doi:10.1186/2041-9139-5-43 Text above from original figure legend. | |||
2041-9139-5-43-2.jpg | 2041-9139-5-43-2.jpg | ||
[[Category:Molecular]][[Category:Cartoon]][[Category:DNA]] |
Revision as of 09:17, 26 March 2015
CRISPR Cas9 interaction with target DNA
he CRISPR/Cas9 complex of Streptococcus pyogenes consists of the Cas9 protein (in gray) and a guide RNA that is a chimera of natural crRNA and tracRNA (in orange).
The targeting sequence at the 5′end of the guide RNA base-pairs with complementary sequences on the target DNA (in blue); the targeting sequence is 20 nucleotides long, but may be shortened to increase specificity (the addition of 1 to 2 unpaired nucleotides at the 5′ end is also tolerated).
The presence of a PAM (protospacer adjacent motif, NGG for Streptococcus pyogenes), located immediately downstream of the 20-nucleotide sequence targeted by the guide RNA, is also essential for target recognition and cleavage.
The PAM sequence does not have a counterpart on the guide RNA. Following recognition of the PAM and base-pairing between the guide RNA and the target,
Cas9 cleaves each of the target DNA strands a few nucleotides upstream of the PAM (red arrowheads). Each strand is cleaved by a different nuclease domain present in Cas9 (HNH and RuvC domains). These domains have been mutated independently to generate Cas9 nickases.
Reference
<pubmed>25699168</pubmed>| Evodevo.
Copright
© 2014 Gilles and Averof; licensee BioMed Central Ltd. This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly credited. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.
Gilles and Averof EvoDevo 2014 5:43 doi:10.1186/2041-9139-5-43 Text above from original figure legend.
2041-9139-5-43-2.jpg
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