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10.1038/cr.2017.76 http://scihub22266oqcxt.onion/10.1038/cr.2017.76 C5518881!5518881!28524166     free     free     free Deprecated: Implicit conversion from float 233.6 to int loses precision in C:\Inetpub\vhosts\kidney.de\httpdocs\pget.php on line 534 Deprecated: Implicit conversion from float 233.6 to int loses precision in C:\Inetpub\vhosts\kidney.de\httpdocs\pget.php on line 534 Deprecated: Implicit conversion from float 233.6 to int loses precision in C:\Inetpub\vhosts\kidney.de\httpdocs\pget.php on line 534       Cell+Res 2017 ; 27 (6): 801-14 Nephropedia Template TP {{tp|p=28524166|t=2017. Homology-mediated end joining-based targeted integration using CRISPR/Cas9 |pdf=|usr=}}{{28524166}} gab.com Text Homology-mediated end joining-based targeted integration using CRISPR/Cas9 JO: Cell Res http://www.kidney.de/pget.php?&tw=1&pmid=28524166 #FOAvip Targeted integration of transgenes can be achieved by strategies based on homologous recombination (HR), microhomology-mediated end joining (MMEJ) or non-homologous end joining (NHEJ). The more generally used HR is inefficient for achieving gene integration in animal embryos and tissues, because it occurs only during cell division, although MMEJ and NHEJ can elevate the efficiency in some systems. Here we devise a homology-mediated end joining (HMEJ)-based strategy, using CRISPR/Cas9-mediated cleavage of both transgene donor vector that contains guide RNA target sites and ?800 bp of homology arms, and the targeted genome. We found no significant improvement of the targeting efficiency by the HMEJ-based method in either mouse embryonic stem cells or the neuroblastoma cell line, N2a, compared to the HR-based method. However, the HMEJ-based method yielded a higher knock-in efficiency in HEK293T cells, primary astrocytes and neurons. More importantly, this approach achieved transgene integration in mouse and monkey embryos, as well as in hepatocytes and neurons in vivo, with an efficiency much greater than HR-, NHEJ- and MMEJ-based strategies. Thus, the HMEJ-based strategy may be useful for a variety of applications, including gene editing to generate animal models and for targeted gene therapies. Twit Text FOAVip Homology-mediated end joining-based targeted integration using CRISPR/Cas9 ????Cell Res http://www.kidney.de/pget.php?&tw=1&pmid=28524166 #FOAvip Twit Text # Free Paper on # # # # # LINK http://www.kidney.de/pget.php?&tw=1&pmid=28524166 #FOAvip English Wikipedia <ref>{{Cite pmid|28524166}}</ref> Homology-mediated end joining-based targeted integration using CRISPR/Cas9 #MMPMID28524166 Yao X; Wang X; Hu X; Liu Z; Liu J; Zhou H; Shen X; Wei Y; Huang Z; Ying W; Wang Y; Nie YH; Zhang CC; Li S; Cheng L; Wang Q; Wu Y; Huang P; Sun Q; Shi L; Yang H Cell Res 2017[Jun]; 27 (6): 801-14 PMID28524166show ga Targeted integration of transgenes can be achieved by strategies based on homologous recombination (HR), microhomology-mediated end joining (MMEJ) or non-homologous end joining (NHEJ). The more generally used HR is inefficient for achieving gene integration in animal embryos and tissues, because it occurs only during cell division, although MMEJ and NHEJ can elevate the efficiency in some systems. Here we devise a homology-mediated end joining (HMEJ)-based strategy, using CRISPR/Cas9-mediated cleavage of both transgene donor vector that contains guide RNA target sites and ?800 bp of homology arms, and the targeted genome. We found no significant improvement of the targeting efficiency by the HMEJ-based method in either mouse embryonic stem cells or the neuroblastoma cell line, N2a, compared to the HR-based method. However, the HMEJ-based method yielded a higher knock-in efficiency in HEK293T cells, primary astrocytes and neurons. More importantly, this approach achieved transgene integration in mouse and monkey embryos, as well as in hepatocytes and neurons in vivo, with an efficiency much greater than HR-, NHEJ- and MMEJ-based strategies. Thus, the HMEJ-based strategy may be useful for a variety of applications, including gene editing to generate animal models and for targeted gene therapies. äHomology-mediated end joining-based targeted integration using CRISPR/(epmc).pdf DeepDyve Pubget Overpricing