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10.1126/sciadv.1701620 http://scihub22266oqcxt.onion/10.1126/sciadv.1701620 C5507636!5507636 !28706995     free     free     free Warning: imagejpeg(C:\Inetpub\vhosts\kidney.de\httpdocs\phplern\28706995 .jpg): Failed to open stream: No such file or directory in C:\Inetpub\vhosts\kidney.de\httpdocs\pget.php on line 117       Sci+Adv 2017 ; 3 (7 ): e1701620 Nephropedia Template TP {{tp|p=28706995 |t=2017. Disabling Cas9 by an anti-CRISPR DNA mimic |pdf=|usr=}}{{28706995 }} gab.com Text Disabling Cas9 by an anti-CRISPR DNA mimic JO: Sci Adv http://www.kidney.de/pget.php?&tw=1&pmid=28706995 #FOAvip CRISPR (clustered regularly interspaced short palindromic repeats)-Cas9 gene editing technology is derived from a microbial adaptive immune system, where bacteriophages are often the intended target. Natural inhibitors of CRISPR-Cas9 enable phages to evade immunity and show promise in controlling Cas9-mediated gene editing in human cells. However, the mechanism of CRISPR-Cas9 inhibition is not known, and the potential applications for Cas9 inhibitor proteins in mammalian cells have not been fully established. We show that the anti-CRISPR protein AcrIIA4 binds only to assembled Cas9-single-guide RNA (sgRNA) complexes and not to Cas9 protein alone. A 3.9 Å resolution cryo-electron microscopy structure of the Cas9-sgRNA-AcrIIA4 complex revealed that the surface of AcrIIA4 is highly acidic and binds with a 1:1 stoichiometry to a region of Cas9 that normally engages the DNA protospacer adjacent motif. Consistent with this binding mode, order-of-addition experiments showed that AcrIIA4 interferes with DNA recognition but has no effect on preformed Cas9-sgRNA-DNA complexes. Timed delivery of AcrIIA4 into human cells as either protein or expression plasmid allows on-target Cas9-mediated gene editing while reducing off-target edits. These results provide a mechanistic understanding of AcrIIA4 function and demonstrate that inhibitors can modulate the extent and outcomes of Cas9-mediated gene editing. Twit Text FOAVip Disabling Cas9 by an anti-CRISPR DNA mimic ????Sci Adv http://www.kidney.de/pget.php?&tw=1&pmid=28706995 #FOAvip Twit Text # Free Paper on # # # # # LINK http://www.kidney.de/pget.php?&tw=1&pmid=28706995 #FOAvip English Wikipedia <ref>{{Cite pmid|28706995 }}</ref> Disabling Cas9 by an anti-CRISPR DNA mimic #MMPMID28706995 Shin J ; Jiang F ; Liu JJ ; Bray NL ; Rauch BJ ; Baik SH ; Nogales E ; Bondy-Denomy J ; Corn JE ; Doudna JA Sci Adv 2017[Jul]; 3 (7 ): e1701620 PMID28706995 show ga CRISPR (clustered regularly interspaced short palindromic repeats)-Cas9 gene editing technology is derived from a microbial adaptive immune system, where bacteriophages are often the intended target. Natural inhibitors of CRISPR-Cas9 enable phages to evade immunity and show promise in controlling Cas9-mediated gene editing in human cells. However, the mechanism of CRISPR-Cas9 inhibition is not known, and the potential applications for Cas9 inhibitor proteins in mammalian cells have not been fully established. We show that the anti-CRISPR protein AcrIIA4 binds only to assembled Cas9-single-guide RNA (sgRNA) complexes and not to Cas9 protein alone. A 3.9 Å resolution cryo-electron microscopy structure of the Cas9-sgRNA-AcrIIA4 complex revealed that the surface of AcrIIA4 is highly acidic and binds with a 1:1 stoichiometry to a region of Cas9 that normally engages the DNA protospacer adjacent motif. Consistent with this binding mode, order-of-addition experiments showed that AcrIIA4 interferes with DNA recognition but has no effect on preformed Cas9-sgRNA-DNA complexes. Timed delivery of AcrIIA4 into human cells as either protein or expression plasmid allows on-target Cas9-mediated gene editing while reducing off-target edits. These results provide a mechanistic understanding of AcrIIA4 function and demonstrate that inhibitors can modulate the extent and outcomes of Cas9-mediated gene editing. |*Clustered Regularly Interspaced Short Palindromic Repeats [MESH] |*Gene Silencing [MESH] |CRISPR-Associated Protein 9/chemistry/*genetics/metabolism [MESH] |CRISPR-Cas Systems [MESH] |Cell Line [MESH] |DNA/*chemistry/*genetics [MESH] |Gene Editing [MESH] |High-Throughput Nucleotide Sequencing [MESH] |Humans [MESH] |Models, Molecular [MESH] |Molecular Conformation [MESH] |RNA, Guide, CRISPR-Cas Systems/genetics [MESH]Disabling Cas9 by an anti-CRISPR DNA mimic (epmc).pdf DeepDyve Pubget Overpricing