5b2o - Revision history

OCA at 09:07, 20 March 2024

OCA — Wed, 20 Mar 2024 09:07:05 GMT

←Older revision		Revision as of 09:07, 20 March 2024
Line 4:		Line 4:
	== Structural highlights ==		== Structural highlights ==
	<table><tr><td colspan='2'>[[5b2o]] is a 4 chain structure with sequence from [https://en.wikipedia.org/wiki/Francisella_tularensis_subsp._novicida Francisella tularensis subsp. novicida], [https://en.wikipedia.org/wiki/Francisella_tularensis_subsp._novicida_U112 Francisella tularensis subsp. novicida U112] and [https://en.wikipedia.org/wiki/Synthetic_construct Synthetic construct]. Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=5B2O OCA]. For a <b>guided tour on the structure components</b> use [https://proteopedia.org/fgij/fg.htm?mol=5B2O FirstGlance]. <br>		<table><tr><td colspan='2'>[[5b2o]] is a 4 chain structure with sequence from [https://en.wikipedia.org/wiki/Francisella_tularensis_subsp._novicida Francisella tularensis subsp. novicida], [https://en.wikipedia.org/wiki/Francisella_tularensis_subsp._novicida_U112 Francisella tularensis subsp. novicida U112] and [https://en.wikipedia.org/wiki/Synthetic_construct Synthetic construct]. Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=5B2O OCA]. For a <b>guided tour on the structure components</b> use [https://proteopedia.org/fgij/fg.htm?mol=5B2O FirstGlance]. <br>
-	</td></tr><tr id='ligand'><td class="sblockLbl"><b>[[Ligand\|Ligands:]]</b></td><td class="sblockDat" id="ligandDat"><scene name='pdbligand=ACT:ACETATE+ION'>ACT</scene>, <scene name='pdbligand=CA:CALCIUM+ION'>CA</scene>, <scene name='pdbligand=CL:CHLORIDE+ION'>CL</scene>, <scene name='pdbligand=EDO:1,2-ETHANEDIOL'>EDO</scene>, <scene name='pdbligand=NA:SODIUM+ION'>NA</scene>, <scene name='pdbligand=ZN:ZINC+ION'>ZN</scene></td></tr>	+	</td></tr><tr id='method'><td class="sblockLbl"><b>[[Empirical_models\|Method:]]</b></td><td class="sblockDat" id="methodDat">X-ray diffraction, [[Resolution\|Resolution]] 1.7Å</td></tr>
		+	<tr id='ligand'><td class="sblockLbl"><b>[[Ligand\|Ligands:]]</b></td><td class="sblockDat" id="ligandDat"><scene name='pdbligand=ACT:ACETATE+ION'>ACT</scene>, <scene name='pdbligand=CA:CALCIUM+ION'>CA</scene>, <scene name='pdbligand=CL:CHLORIDE+ION'>CL</scene>, <scene name='pdbligand=EDO:1,2-ETHANEDIOL'>EDO</scene>, <scene name='pdbligand=NA:SODIUM+ION'>NA</scene>, <scene name='pdbligand=ZN:ZINC+ION'>ZN</scene></td></tr>
	<tr id='resources'><td class="sblockLbl"><b>Resources:</b></td><td class="sblockDat"><span class='plainlinks'>[https://proteopedia.org/fgij/fg.htm?mol=5b2o FirstGlance], [http://oca.weizmann.ac.il/oca-bin/ocaids?id=5b2o OCA], [https://pdbe.org/5b2o PDBe], [https://www.rcsb.org/pdb/explore.do?structureId=5b2o RCSB], [https://www.ebi.ac.uk/pdbsum/5b2o PDBsum], [https://prosat.h-its.org/prosat/prosatexe?pdbcode=5b2o ProSAT]</span></td></tr>		<tr id='resources'><td class="sblockLbl"><b>Resources:</b></td><td class="sblockDat"><span class='plainlinks'>[https://proteopedia.org/fgij/fg.htm?mol=5b2o FirstGlance], [http://oca.weizmann.ac.il/oca-bin/ocaids?id=5b2o OCA], [https://pdbe.org/5b2o PDBe], [https://www.rcsb.org/pdb/explore.do?structureId=5b2o RCSB], [https://www.ebi.ac.uk/pdbsum/5b2o PDBsum], [https://prosat.h-its.org/prosat/prosatexe?pdbcode=5b2o ProSAT]</span></td></tr>
	</table>		</table>
	== Function ==		== Function ==
	[https://www.uniprot.org/uniprot/CAS9_FRATN CAS9_FRATN] CRISPR (clustered regularly interspaced short palindromic repeat) is an adaptive immune system that provides protection against mobile genetic elements (viruses, transposable elements and conjugative plasmids). CRISPR clusters contain spacers, sequences complementary to antecedent mobile elements, and target invading nucleic acids. CRISPR clusters are transcribed and processed into CRISPR RNA (crRNA). In type II CRISPR systems correct processing of pre-crRNA requires a trans-encoded small RNA (tracrRNA), endogenous ribonuclease 3 (rnc) and this protein. The tracrRNA serves as a guide for ribonuclease 3-aided processing of pre-crRNA. Subsequently Cas9/crRNA/tracrRNA endonucleolytically cleaves linear or circular dsDNA target complementary to the spacer; Cas9 is inactive in the absence of the 2 guide RNAs (gRNA). Cas9 recognizes a short motif in the CRISPR repeat sequences (the PAM or protospacer adjacent motif) to help distinguish self versus nonself, as targets within the bacterial CRISPR locus do not have PAMs. PAM recognition is also required for catalytic activity (By similarity). Cuts target DNA when Cas9 and gRNAs are mixed.<ref>PMID:24270795</ref>		[https://www.uniprot.org/uniprot/CAS9_FRATN CAS9_FRATN] CRISPR (clustered regularly interspaced short palindromic repeat) is an adaptive immune system that provides protection against mobile genetic elements (viruses, transposable elements and conjugative plasmids). CRISPR clusters contain spacers, sequences complementary to antecedent mobile elements, and target invading nucleic acids. CRISPR clusters are transcribed and processed into CRISPR RNA (crRNA). In type II CRISPR systems correct processing of pre-crRNA requires a trans-encoded small RNA (tracrRNA), endogenous ribonuclease 3 (rnc) and this protein. The tracrRNA serves as a guide for ribonuclease 3-aided processing of pre-crRNA. Subsequently Cas9/crRNA/tracrRNA endonucleolytically cleaves linear or circular dsDNA target complementary to the spacer; Cas9 is inactive in the absence of the 2 guide RNAs (gRNA). Cas9 recognizes a short motif in the CRISPR repeat sequences (the PAM or protospacer adjacent motif) to help distinguish self versus nonself, as targets within the bacterial CRISPR locus do not have PAMs. PAM recognition is also required for catalytic activity (By similarity). Cuts target DNA when Cas9 and gRNAs are mixed.<ref>PMID:24270795</ref>
-	<div style="background-color:#fffaf0;">
-	== Publication Abstract from PubMed ==
-	The RNA-guided endonuclease Cas9 cleaves double-stranded DNA targets complementary to the guide RNA and has been applied to programmable genome editing. Cas9-mediated cleavage requires a protospacer adjacent motif (PAM) juxtaposed with the DNA target sequence, thus constricting the range of targetable sites. Here, we report the 1.7 A resolution crystal structures of Cas9 from Francisella novicida (FnCas9), one of the largest Cas9 orthologs, in complex with a guide RNA and its PAM-containing DNA targets. A structural comparison of FnCas9 with other Cas9 orthologs revealed striking conserved and divergent features among distantly related CRISPR-Cas9 systems. We found that FnCas9 recognizes the 5'-NGG-3' PAM, and used the structural information to create a variant that can recognize the more relaxed 5'-YG-3' PAM. Furthermore, we demonstrated that the FnCas9-ribonucleoprotein complex can be microinjected into mouse zygotes to edit endogenous sites with the 5'-YG-3' PAM, thus expanding the target space of the CRISPR-Cas9 toolbox.
-
-	Structure and Engineering of Francisella novicida Cas9.,Hirano H, Gootenberg JS, Horii T, Abudayyeh OO, Kimura M, Hsu PD, Nakane T, Ishitani R, Hatada I, Zhang F, Nishimasu H, Nureki O Cell. 2016 Feb 25;164(5):950-61. doi: 10.1016/j.cell.2016.01.039. Epub 2016 Feb, 11. PMID:26875867<ref>PMID:26875867</ref>
-
-	From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine.<br>
-	</div>
-	<div class="pdbe-citations 5b2o" style="background-color:#fffaf0;"></div>

	==See Also==		==See Also==

OCA at 06:36, 31 May 2023

OCA — Wed, 31 May 2023 06:36:51 GMT

←Older revision		Revision as of 06:36, 31 May 2023
Line 3:		Line 3:
	<StructureSection load='5b2o' size='340' side='right'caption='[[5b2o]], [[Resolution\|resolution]] 1.70Å' scene=''>		<StructureSection load='5b2o' size='340' side='right'caption='[[5b2o]], [[Resolution\|resolution]] 1.70Å' scene=''>
	== Structural highlights ==		== Structural highlights ==
-	<table><tr><td colspan='2'>[[5b2o]] is a 4 chain structure with sequence from [~~http~~://en.wikipedia.org/wiki/~~Fratn Fratn~~]. Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=5B2O OCA]. For a <b>guided tour on the structure components</b> use [~~http~~://~~oca~~.~~weizmann.ac.il/oca-docs~~/fgij/fg.htm?mol=5B2O FirstGlance]. <br>	+	<table><tr><td colspan='2'>[[5b2o]] is a 4 chain structure with sequence from [https://en.wikipedia.org/wiki/Francisella_tularensis_subsp._novicida Francisella tularensis subsp. novicida], [https://en.wikipedia.org/wiki/Francisella_tularensis_subsp._novicida_U112 Francisella tularensis subsp. novicida U112] and [https://en.wikipedia.org/wiki/Synthetic_construct Synthetic construct]. Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=5B2O OCA]. For a <b>guided tour on the structure components</b> use [https://proteopedia.org/fgij/fg.htm?mol=5B2O FirstGlance]. <br>
-	</td></tr><tr id='ligand'><td class="sblockLbl"><b>[[Ligand\|Ligands:]]</b></td><td class="sblockDat"><scene name='pdbligand=ACT:ACETATE+ION'>ACT</scene>, <scene name='pdbligand=CA:CALCIUM+ION'>CA</scene>, <scene name='pdbligand=CL:CHLORIDE+ION'>CL</scene>, <scene name='pdbligand=EDO:1,2-ETHANEDIOL'>EDO</scene>, <scene name='pdbligand=NA:SODIUM+ION'>NA</scene>, <scene name='pdbligand=ZN:ZINC+ION'>ZN</scene>~~</td></tr>~~	+	</td></tr><tr id='ligand'><td class="sblockLbl"><b>[[Ligand\|Ligands:]]</b></td><td class="sblockDat" id="ligandDat"><scene name='pdbligand=ACT:ACETATE+ION'>ACT</scene>, <scene name='pdbligand=CA:CALCIUM+ION'>CA</scene>, <scene name='pdbligand=CL:CHLORIDE+ION'>CL</scene>, <scene name='pdbligand=EDO:1,2-ETHANEDIOL'>EDO</scene>, <scene name='pdbligand=NA:SODIUM+ION'>NA</scene>, <scene name='pdbligand=ZN:ZINC+ION'>ZN</scene></td></tr>
-	~~<tr id='NonStdRes'><td class="sblockLbl"><b>[[Non-Standard_Residue\|NonStd Res:]]</b></td><td class="sblockDat"><scene name='pdbligand=DU:2-DEOXYURIDINE-5-MONOPHOSPHATE'>DU</scene></td></tr>~~	+	<tr id='resources'><td class="sblockLbl"><b>Resources:</b></td><td class="sblockDat"><span class='plainlinks'>[https://proteopedia.org/fgij/fg.htm?mol=5b2o FirstGlance], [http://oca.weizmann.ac.il/oca-bin/ocaids?id=5b2o OCA], [https://pdbe.org/5b2o PDBe], [https://www.rcsb.org/pdb/explore.do?structureId=5b2o RCSB], [https://www.ebi.ac.uk/pdbsum/5b2o PDBsum], [https://prosat.h-its.org/prosat/prosatexe?pdbcode=5b2o ProSAT]</span></td></tr>
-	~~<tr id='related'><td class="sblockLbl"><b>[[Related_structure\|Related:]]</b></td><td class="sblockDat">[[5b2p\|5b2p]], [[5b2q\|5b2q]]</td></tr>~~	+
-	~~<tr id='gene'><td class="sblockLbl"><b>[[Gene\|Gene:]]</b></td><td class="sblockDat">cas9, FTN_0757 ([http://www.ncbi.nlm.nih.gov/Taxonomy/Browser/wwwtax.cgi?mode=Info&srchmode=5&id=401614 FRATN])~~</td></tr>	+
-	<tr id='resources'><td class="sblockLbl"><b>Resources:</b></td><td class="sblockDat"><span class='plainlinks'>[~~http~~://~~oca~~.~~weizmann.ac.il/oca-docs~~/fgij/fg.htm?mol=5b2o FirstGlance], [http://oca.weizmann.ac.il/oca-bin/ocaids?id=5b2o OCA], [~~http~~://pdbe.org/5b2o PDBe], [~~http~~://www.rcsb.org/pdb/explore.do?structureId=5b2o RCSB], [~~http~~://www.ebi.ac.uk/pdbsum/5b2o PDBsum], [~~http~~://prosat.h-its.org/prosat/prosatexe?pdbcode=5b2o ProSAT]</span></td></tr>	+
	</table>		</table>
	== Function ==		== Function ==
-	[~~[http~~://www.uniprot.org/uniprot/CAS9_FRATN CAS9_FRATN]] CRISPR (clustered regularly interspaced short palindromic repeat) is an adaptive immune system that provides protection against mobile genetic elements (viruses, transposable elements and conjugative plasmids). CRISPR clusters contain spacers, sequences complementary to antecedent mobile elements, and target invading nucleic acids. CRISPR clusters are transcribed and processed into CRISPR RNA (crRNA). In type II CRISPR systems correct processing of pre-crRNA requires a trans-encoded small RNA (tracrRNA), endogenous ribonuclease 3 (rnc) and this protein. The tracrRNA serves as a guide for ribonuclease 3-aided processing of pre-crRNA. Subsequently Cas9/crRNA/tracrRNA endonucleolytically cleaves linear or circular dsDNA target complementary to the spacer; Cas9 is inactive in the absence of the 2 guide RNAs (gRNA). Cas9 recognizes a short motif in the CRISPR repeat sequences (the PAM or protospacer adjacent motif) to help distinguish self versus nonself, as targets within the bacterial CRISPR locus do not have PAMs. PAM recognition is also required for catalytic activity (By similarity). Cuts target DNA when Cas9 and gRNAs are mixed.<ref>PMID:24270795</ref>	+	[https://www.uniprot.org/uniprot/CAS9_FRATN CAS9_FRATN] CRISPR (clustered regularly interspaced short palindromic repeat) is an adaptive immune system that provides protection against mobile genetic elements (viruses, transposable elements and conjugative plasmids). CRISPR clusters contain spacers, sequences complementary to antecedent mobile elements, and target invading nucleic acids. CRISPR clusters are transcribed and processed into CRISPR RNA (crRNA). In type II CRISPR systems correct processing of pre-crRNA requires a trans-encoded small RNA (tracrRNA), endogenous ribonuclease 3 (rnc) and this protein. The tracrRNA serves as a guide for ribonuclease 3-aided processing of pre-crRNA. Subsequently Cas9/crRNA/tracrRNA endonucleolytically cleaves linear or circular dsDNA target complementary to the spacer; Cas9 is inactive in the absence of the 2 guide RNAs (gRNA). Cas9 recognizes a short motif in the CRISPR repeat sequences (the PAM or protospacer adjacent motif) to help distinguish self versus nonself, as targets within the bacterial CRISPR locus do not have PAMs. PAM recognition is also required for catalytic activity (By similarity). Cuts target DNA when Cas9 and gRNAs are mixed.<ref>PMID:24270795</ref>
	<div style="background-color:#fffaf0;">		<div style="background-color:#fffaf0;">
	== Publication Abstract from PubMed ==		== Publication Abstract from PubMed ==
Line 28:		Line 25:
	__TOC__		__TOC__
	</StructureSection>		</StructureSection>
-	[[Category: ~~Fratn~~]]	+	[[Category: Francisella tularensis subsp. novicida]]
		+	[[Category: Francisella tularensis subsp. novicida U112]]
	[[Category: Large Structures]]		[[Category: Large Structures]]
-	[[Category: Hirano, H]]	+	[[Category: Synthetic construct]]
-	[[Category: Ishitani, R]]	+	[[Category: Hirano H]]
-	[[Category: Nakane, T]]	+	[[Category: Ishitani R]]
-	[[Category: Nishimasu, H]]	+	[[Category: Nakane T]]
-	[[Category: Nureki, O]]	+	[[Category: Nishimasu H]]
-	~~[[Category: Crispr-cas9]]~~	+	[[Category: Nureki O]]
-	~~[[Category: Genome engineering]]~~	+
-	~~[[Category: Hydrolase-rna-dna complex~~]]	+

OCA at 08:45, 11 March 2020

OCA — Wed, 11 Mar 2020 08:45:22 GMT

←Older revision		Revision as of 08:45, 11 March 2020
Line 1:		Line 1:

	==Crystal structure of Francisella novicida Cas9 in complex with sgRNA and target DNA (TGG PAM)==		==Crystal structure of Francisella novicida Cas9 in complex with sgRNA and target DNA (TGG PAM)==
-	<StructureSection load='5b2o' size='340' side='right' caption='[[5b2o]], [[Resolution\|resolution]] 1.70Å' scene=''>	+	<StructureSection load='5b2o' size='340' side='right'caption='[[5b2o]], [[Resolution\|resolution]] 1.70Å' scene=''>
	== Structural highlights ==		== Structural highlights ==
-	<table><tr><td colspan='2'>[[5b2o]] is a 4 chain structure. Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=5B2O OCA]. For a <b>guided tour on the structure components</b> use [http://oca.weizmann.ac.il/oca-docs/fgij/fg.htm?mol=5B2O FirstGlance]. <br>	+	<table><tr><td colspan='2'>[[5b2o]] is a 4 chain structure with sequence from [http://en.wikipedia.org/wiki/Fratn Fratn]. Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=5B2O OCA]. For a <b>guided tour on the structure components</b> use [http://oca.weizmann.ac.il/oca-docs/fgij/fg.htm?mol=5B2O FirstGlance]. <br>
	</td></tr><tr id='ligand'><td class="sblockLbl"><b>[[Ligand\|Ligands:]]</b></td><td class="sblockDat"><scene name='pdbligand=ACT:ACETATE+ION'>ACT</scene>, <scene name='pdbligand=CA:CALCIUM+ION'>CA</scene>, <scene name='pdbligand=CL:CHLORIDE+ION'>CL</scene>, <scene name='pdbligand=EDO:1,2-ETHANEDIOL'>EDO</scene>, <scene name='pdbligand=NA:SODIUM+ION'>NA</scene>, <scene name='pdbligand=ZN:ZINC+ION'>ZN</scene></td></tr>		</td></tr><tr id='ligand'><td class="sblockLbl"><b>[[Ligand\|Ligands:]]</b></td><td class="sblockDat"><scene name='pdbligand=ACT:ACETATE+ION'>ACT</scene>, <scene name='pdbligand=CA:CALCIUM+ION'>CA</scene>, <scene name='pdbligand=CL:CHLORIDE+ION'>CL</scene>, <scene name='pdbligand=EDO:1,2-ETHANEDIOL'>EDO</scene>, <scene name='pdbligand=NA:SODIUM+ION'>NA</scene>, <scene name='pdbligand=ZN:ZINC+ION'>ZN</scene></td></tr>
	<tr id='NonStdRes'><td class="sblockLbl"><b>[[Non-Standard_Residue\|NonStd Res:]]</b></td><td class="sblockDat"><scene name='pdbligand=DU:2-DEOXYURIDINE-5-MONOPHOSPHATE'>DU</scene></td></tr>		<tr id='NonStdRes'><td class="sblockLbl"><b>[[Non-Standard_Residue\|NonStd Res:]]</b></td><td class="sblockDat"><scene name='pdbligand=DU:2-DEOXYURIDINE-5-MONOPHOSPHATE'>DU</scene></td></tr>
	<tr id='related'><td class="sblockLbl"><b>[[Related_structure\|Related:]]</b></td><td class="sblockDat">[[5b2p\|5b2p]], [[5b2q\|5b2q]]</td></tr>		<tr id='related'><td class="sblockLbl"><b>[[Related_structure\|Related:]]</b></td><td class="sblockDat">[[5b2p\|5b2p]], [[5b2q\|5b2q]]</td></tr>
-	<tr id='resources'><td class="sblockLbl"><b>Resources:</b></td><td class="sblockDat"><span class='plainlinks'>[http://oca.weizmann.ac.il/oca-docs/fgij/fg.htm?mol=5b2o FirstGlance], [http://oca.weizmann.ac.il/oca-bin/ocaids?id=5b2o OCA], [http://pdbe.org/5b2o PDBe], [http://www.rcsb.org/pdb/explore.do?structureId=5b2o RCSB], [http://www.ebi.ac.uk/pdbsum/5b2o PDBsum]</span></td></tr>	+	<tr id='gene'><td class="sblockLbl"><b>[[Gene\|Gene:]]</b></td><td class="sblockDat">cas9, FTN_0757 ([http://www.ncbi.nlm.nih.gov/Taxonomy/Browser/wwwtax.cgi?mode=Info&srchmode=5&id=401614 FRATN])</td></tr>
		+	<tr id='resources'><td class="sblockLbl"><b>Resources:</b></td><td class="sblockDat"><span class='plainlinks'>[http://oca.weizmann.ac.il/oca-docs/fgij/fg.htm?mol=5b2o FirstGlance], [http://oca.weizmann.ac.il/oca-bin/ocaids?id=5b2o OCA], [http://pdbe.org/5b2o PDBe], [http://www.rcsb.org/pdb/explore.do?structureId=5b2o RCSB], [http://www.ebi.ac.uk/pdbsum/5b2o PDBsum], [http://prosat.h-its.org/prosat/prosatexe?pdbcode=5b2o ProSAT]</span></td></tr>
	</table>		</table>
	== Function ==		== Function ==
Line 20:		Line 21:
	</div>		</div>
	<div class="pdbe-citations 5b2o" style="background-color:#fffaf0;"></div>		<div class="pdbe-citations 5b2o" style="background-color:#fffaf0;"></div>
		+
		+	==See Also==
		+	*[[Endonuclease 3D structures\|Endonuclease 3D structures]]
	== References ==		== References ==
	<references/>		<references/>
	__TOC__		__TOC__
	</StructureSection>		</StructureSection>
		+	[[Category: Fratn]]
		+	[[Category: Large Structures]]
	[[Category: Hirano, H]]		[[Category: Hirano, H]]
	[[Category: Ishitani, R]]		[[Category: Ishitani, R]]

OCA at 14:58, 2 March 2016

OCA — Wed, 02 Mar 2016 14:58:31 GMT

←Older revision		Revision as of 14:58, 2 March 2016
Line 1:		Line 1:
-	'''Unreleased structure'''

-	~~The entry~~ 5b2o is ~~ON HOLD~~	+	==Crystal structure of Francisella novicida Cas9 in complex with sgRNA and target DNA (TGG PAM)==
		+	<StructureSection load='5b2o' size='340' side='right' caption='[[5b2o]], [[Resolution\|resolution]] 1.70Å' scene=''>
		+	== Structural highlights ==
		+	<table><tr><td colspan='2'>[[5b2o]] is a 4 chain structure. Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=5B2O OCA]. For a <b>guided tour on the structure components</b> use [http://oca.weizmann.ac.il/oca-docs/fgij/fg.htm?mol=5B2O FirstGlance]. <br>
		+	</td></tr><tr id='ligand'><td class="sblockLbl"><b>[[Ligand\|Ligands:]]</b></td><td class="sblockDat"><scene name='pdbligand=ACT:ACETATE+ION'>ACT</scene>, <scene name='pdbligand=CA:CALCIUM+ION'>CA</scene>, <scene name='pdbligand=CL:CHLORIDE+ION'>CL</scene>, <scene name='pdbligand=EDO:1,2-ETHANEDIOL'>EDO</scene>, <scene name='pdbligand=NA:SODIUM+ION'>NA</scene>, <scene name='pdbligand=ZN:ZINC+ION'>ZN</scene></td></tr>
		+	<tr id='NonStdRes'><td class="sblockLbl"><b>[[Non-Standard_Residue\|NonStd Res:]]</b></td><td class="sblockDat"><scene name='pdbligand=DU:2-DEOXYURIDINE-5-MONOPHOSPHATE'>DU</scene></td></tr>
		+	<tr id='related'><td class="sblockLbl"><b>[[Related_structure\|Related:]]</b></td><td class="sblockDat">[[5b2p\|5b2p]], [[5b2q\|5b2q]]</td></tr>
		+	<tr id='resources'><td class="sblockLbl"><b>Resources:</b></td><td class="sblockDat"><span class='plainlinks'>[http://oca.weizmann.ac.il/oca-docs/fgij/fg.htm?mol=5b2o FirstGlance], [http://oca.weizmann.ac.il/oca-bin/ocaids?id=5b2o OCA], [http://pdbe.org/5b2o PDBe], [http://www.rcsb.org/pdb/explore.do?structureId=5b2o RCSB], [http://www.ebi.ac.uk/pdbsum/5b2o PDBsum]</span></td></tr>
		+	</table>
		+	== Function ==
		+	[[http://www.uniprot.org/uniprot/CAS9_FRATN CAS9_FRATN]] CRISPR (clustered regularly interspaced short palindromic repeat) is an adaptive immune system that provides protection against mobile genetic elements (viruses, transposable elements and conjugative plasmids). CRISPR clusters contain spacers, sequences complementary to antecedent mobile elements, and target invading nucleic acids. CRISPR clusters are transcribed and processed into CRISPR RNA (crRNA). In type II CRISPR systems correct processing of pre-crRNA requires a trans-encoded small RNA (tracrRNA), endogenous ribonuclease 3 (rnc) and this protein. The tracrRNA serves as a guide for ribonuclease 3-aided processing of pre-crRNA. Subsequently Cas9/crRNA/tracrRNA endonucleolytically cleaves linear or circular dsDNA target complementary to the spacer; Cas9 is inactive in the absence of the 2 guide RNAs (gRNA). Cas9 recognizes a short motif in the CRISPR repeat sequences (the PAM or protospacer adjacent motif) to help distinguish self versus nonself, as targets within the bacterial CRISPR locus do not have PAMs. PAM recognition is also required for catalytic activity (By similarity). Cuts target DNA when Cas9 and gRNAs are mixed.<ref>PMID:24270795</ref>
		+	<div style="background-color:#fffaf0;">
		+	== Publication Abstract from PubMed ==
		+	The RNA-guided endonuclease Cas9 cleaves double-stranded DNA targets complementary to the guide RNA and has been applied to programmable genome editing. Cas9-mediated cleavage requires a protospacer adjacent motif (PAM) juxtaposed with the DNA target sequence, thus constricting the range of targetable sites. Here, we report the 1.7 A resolution crystal structures of Cas9 from Francisella novicida (FnCas9), one of the largest Cas9 orthologs, in complex with a guide RNA and its PAM-containing DNA targets. A structural comparison of FnCas9 with other Cas9 orthologs revealed striking conserved and divergent features among distantly related CRISPR-Cas9 systems. We found that FnCas9 recognizes the 5'-NGG-3' PAM, and used the structural information to create a variant that can recognize the more relaxed 5'-YG-3' PAM. Furthermore, we demonstrated that the FnCas9-ribonucleoprotein complex can be microinjected into mouse zygotes to edit endogenous sites with the 5'-YG-3' PAM, thus expanding the target space of the CRISPR-Cas9 toolbox.

-	~~Authors: Hirano~~, H., ~~Nishimasu~~, H., ~~Nakane~~, T., Ishitani, R., ~~Nureki~~, O.	+	Structure and Engineering of Francisella novicida Cas9.,Hirano H, Gootenberg JS, Horii T, Abudayyeh OO, Kimura M, Hsu PD, Nakane T, Ishitani R, Hatada I, Zhang F, Nishimasu H, Nureki O Cell. 2016 Feb 25;164(5):950-61. doi: 10.1016/j.cell.2016.01.039. Epub 2016 Feb, 11. PMID:26875867<ref>PMID:26875867</ref>

-	~~Description: Crystal structure~~ of ~~Francisella novicida Cas9 in complex with sgRNA and target DNA (TGG PAM)~~	+	From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine.<br>
-	~~[[Category: Unreleased Structures]]~~	+	</div>
-	~~[[Category~~: ~~Nureki, O]]~~	+	<div class="pdbe-citations 5b2o" style="background-color:#fffaf0;"></div>
-	~~[[Category: Nishimasu, H]]~~	+	== References ==
-	~~[[Category: Ishitani, R]]~~	+	<references/>
		+	__TOC__
		+	</StructureSection>
	[[Category: Hirano, H]]		[[Category: Hirano, H]]
		+	[[Category: Ishitani, R]]
	[[Category: Nakane, T]]		[[Category: Nakane, T]]
		+	[[Category: Nishimasu, H]]
		+	[[Category: Nureki, O]]
		+	[[Category: Crispr-cas9]]
		+	[[Category: Genome engineering]]
		+	[[Category: Hydrolase-rna-dna complex]]

OCA at 12:22, 26 February 2016

OCA — Fri, 26 Feb 2016 12:22:15 GMT

←Older revision		Revision as of 12:22, 26 February 2016
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	The entry 5b2o is ON HOLD		The entry 5b2o is ON HOLD

-	Authors: Hirano, H., Nishimasu, H., Nakane, T, Ishitani, R., Nureki, O.	+	Authors: Hirano, H., Nishimasu, H., Nakane, T., Ishitani, R., Nureki, O.

	Description: Crystal structure of Francisella novicida Cas9 in complex with sgRNA and target DNA (TGG PAM)		Description: Crystal structure of Francisella novicida Cas9 in complex with sgRNA and target DNA (TGG PAM)
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	[[Category: Nureki, O]]		[[Category: Nureki, O]]
	[[Category: Nishimasu, H]]		[[Category: Nishimasu, H]]
		+	[[Category: Ishitani, R]]
	[[Category: Hirano, H]]		[[Category: Hirano, H]]
-	[[Category: Nakane, T~~, Ishitani, R~~]]	+	[[Category: Nakane, T]]

OCA: Protected "5b2o" [edit=sysop:move=sysop]

OCA — Wed, 10 Feb 2016 11:35:36 GMT

Protected "5b2o" [edit=sysop:move=sysop]

←Older revision

Revision as of 11:35, 10 February 2016

OCA: New page: '''Unreleased structure''' The entry 5b2o is ON HOLD Authors: Hirano, H., Nishimasu, H., Nakane, T, Ishitani, R., Nureki, O. Description: Crystal structure of Francisella novicida Cas9...

OCA — Wed, 10 Feb 2016 11:35:35 GMT

New page: '''Unreleased structure''' The entry 5b2o is ON HOLD Authors: Hirano, H., Nishimasu, H., Nakane, T, Ishitani, R., Nureki, O. Description: Crystal structure of Francisella novicida Cas9...

New page

'''Unreleased structure'''

The entry 5b2o is ON HOLD

Authors: Hirano, H., Nishimasu, H., Nakane, T, Ishitani, R., Nureki, O.

Description: Crystal structure of Francisella novicida Cas9 in complex with sgRNA and target DNA (TGG PAM)
[[Category: Unreleased Structures]]
[[Category: Nureki, O]]
[[Category: Nishimasu, H]]
[[Category: Hirano, H]]
[[Category: Nakane, T, Ishitani, R]]