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10.1038/s41467-018-05245-x http://scihub22266oqcxt.onion/10.1038/s41467-018-05245-x C6050341!6050341!30018371     free     free     free Deprecated: Implicit conversion from float 231.6 to int loses precision in C:\Inetpub\vhosts\kidney.de\httpdocs\pget.php on line 534 Deprecated: Implicit conversion from float 231.6 to int loses precision in C:\Inetpub\vhosts\kidney.de\httpdocs\pget.php on line 534 Deprecated: Implicit conversion from float 231.6 to int loses precision in C:\Inetpub\vhosts\kidney.de\httpdocs\pget.php on line 534       Nat+Commun 2018 ; 9 (ä): ä Nephropedia Template TP {{tp|p=30018371|t=2018. Direct observation of DNA target searching and cleavage by CRISPR-Cas12a |pdf=|usr=}}{{30018371}} gab.com Text Direct observation of DNA target searching and cleavage by CRISPR-Cas12a JO: Nat Commun http://www.kidney.de/pget.php?&tw=1&pmid=30018371 #FOAvip Cas12a (also called Cpf1) is a representative type V-A CRISPR effector RNA-guided DNA endonuclease, which provides an alternative to type II CRISPR?Cas9 for genome editing. Previous studies have revealed that Cas12a has unique features distinct from Cas9, but the detailed mechanisms of target searching and DNA cleavage by Cas12a are still unclear. Here, we directly observe this entire process by using single-molecule fluorescence assays to study Cas12a from Acidaminococcus sp. (AsCas12a). We determine that AsCas12a ribonucleoproteins search for their on-target site by a one-dimensional diffusion along elongated DNA molecules and induce cleavage in the two DNA strands in a well-defined order, beginning with the non-target strand. Furthermore, the protospacer-adjacent motif (PAM) for AsCas12a makes only a limited contribution of DNA unwinding during R-loop formation and shows a negligible role in the process of DNA cleavage, in contrast to the Cas9 PAM. Twit Text FOAVip Direct observation of DNA target searching and cleavage by CRISPR-Cas12a ????Nat Commun http://www.kidney.de/pget.php?&tw=1&pmid=30018371 #FOAvip Twit Text # Free Paper on # # # # # LINK http://www.kidney.de/pget.php?&tw=1&pmid=30018371 #FOAvip English Wikipedia <ref>{{Cite pmid|30018371}}</ref> Direct observation of DNA target searching and cleavage by CRISPR-Cas12a #MMPMID30018371 Jeon Y; Choi YH; Jang Y; Yu J; Goo J; Lee G; Jeong YK; Lee SH; Kim IS; Kim JS; Jeong C; Lee S; Bae S Nat Commun 2018[]; 9 (ä): ä PMID30018371show ga Cas12a (also called Cpf1) is a representative type V-A CRISPR effector RNA-guided DNA endonuclease, which provides an alternative to type II CRISPR?Cas9 for genome editing. Previous studies have revealed that Cas12a has unique features distinct from Cas9, but the detailed mechanisms of target searching and DNA cleavage by Cas12a are still unclear. Here, we directly observe this entire process by using single-molecule fluorescence assays to study Cas12a from Acidaminococcus sp. (AsCas12a). We determine that AsCas12a ribonucleoproteins search for their on-target site by a one-dimensional diffusion along elongated DNA molecules and induce cleavage in the two DNA strands in a well-defined order, beginning with the non-target strand. Furthermore, the protospacer-adjacent motif (PAM) for AsCas12a makes only a limited contribution of DNA unwinding during R-loop formation and shows a negligible role in the process of DNA cleavage, in contrast to the Cas9 PAM. äDirect observation of DNA target searching and cleavage by CRISPR-Cas1(epmc).pdf DeepDyve Pubget Overpricing