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10.1186/s13059-015-0817-8 http://scihub22266oqcxt.onion/10.1186/s13059-015-0817-8 C4657203!4657203!26596280     free     free     free Warning: file_get_contents(https://eutils.ncbi.nlm.nih.gov/entrez/eutils/elink.fcgi?dbfrom=pubmed&id=26596280&cmd=llinks): Failed to open stream: HTTP request failed! 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Characterization of Staphylococcus aureus Cas9: a smaller Cas9 for all-in-one adeno-associated virus delivery and paired nickase applications |pdf=|usr=}}{{26596280}} gab.com Text Characterization of Staphylococcus aureus Cas9: a smaller Cas9 for all-in-one adeno-associated virus delivery and paired nickase applications JO: Genome Biol http://www.kidney.de/pget.php?&tw=1&pmid=26596280 #FOAvip Background: CRISPR-Cas systems have been broadly embraced as effective tools for genome engineering applications, with most studies to date utilizing the Streptococcus pyogenes Cas9. Here we characterize and manipulate the smaller, 1053 amino acid nuclease Staphylococcus aureus Cas9. Results: We find that the S. aureus Cas9 recognizes an NNGRRT protospacer adjacent motif (PAM) and cleaves target DNA at high efficiency with a variety of guide RNA (gRNA) spacer lengths. When directed against genomic targets with mutually permissive NGGRRT PAMs, the S. pyogenes Cas9 and S. aureus Cas9 yield indels at comparable rates. We additionally show D10A and N580A paired nickase activity with S. aureus Cas9, and we further package it with two gRNAs in a single functional adeno-associated virus (AAV) vector. Finally, we assess comparative S. pyogenes and S. aureus Cas9 specificity using GUIDE-seq. Conclusion: Our results reveal an S. aureus Cas9 that is effective for a variety of genome engineering purposes, including paired nickase approaches and all-in-one delivery of Cas9 and multiple gRNA expression cassettes with AAV vectors. Electronic supplementary material: The online version of this article (doi:10.1186/s13059-015-0817-8) contains supplementary material, which is available to authorized users. Twit Text FOAVip Characterization of Staphylococcus aureus Cas9: a smaller Cas9 for all-in-one adeno-associated virus delivery and paired nickase applications ????Genome Biol http://www.kidney.de/pget.php?&tw=1&pmid=26596280 #FOAvip Twit Text # Free Paper on # # # # # LINK http://www.kidney.de/pget.php?&tw=1&pmid=26596280 #FOAvip English Wikipedia <ref>{{Cite pmid|26596280}}</ref> Characterization of Staphylococcus aureus Cas9: a smaller Cas9 for all-in-one adeno-associated virus delivery and paired nickase applications #MMPMID26596280 Friedland AE; Baral R; Singhal P; Loveluck K; Shen S; Sanchez M; Marco E; Gotta GM; Maeder ML; Kennedy EM; Kornepati AVR; Sousa A; Collins MA; Jayaram H; Cullen BR; Bumcrot D Genome Biol 2015[]; 16 (ä): ä PMID26596280show ga Background: CRISPR-Cas systems have been broadly embraced as effective tools for genome engineering applications, with most studies to date utilizing the Streptococcus pyogenes Cas9. Here we characterize and manipulate the smaller, 1053 amino acid nuclease Staphylococcus aureus Cas9. Results: We find that the S. aureus Cas9 recognizes an NNGRRT protospacer adjacent motif (PAM) and cleaves target DNA at high efficiency with a variety of guide RNA (gRNA) spacer lengths. When directed against genomic targets with mutually permissive NGGRRT PAMs, the S. pyogenes Cas9 and S. aureus Cas9 yield indels at comparable rates. We additionally show D10A and N580A paired nickase activity with S. aureus Cas9, and we further package it with two gRNAs in a single functional adeno-associated virus (AAV) vector. Finally, we assess comparative S. pyogenes and S. aureus Cas9 specificity using GUIDE-seq. Conclusion: Our results reveal an S. aureus Cas9 that is effective for a variety of genome engineering purposes, including paired nickase approaches and all-in-one delivery of Cas9 and multiple gRNA expression cassettes with AAV vectors. Electronic supplementary material: The online version of this article (doi:10.1186/s13059-015-0817-8) contains supplementary material, which is available to authorized users. äCharacterization of Staphylococcus aureus Cas9: a smaller Cas9 for all(epmc).pdf DeepDyve Pubget Overpricing