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10.1007/s13238-015-0153-5 http://scihub22266oqcxt.onion/10.1007/s13238-015-0153-5 25894090!4417674!25894090     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       Protein+Cell 2015 ; 6 (5): 363-372 Nephropedia Template TP {{tp|p=25894090|t=2015. CRISPR/Cas9-mediated gene editing in human tripronuclear zygotes |pdf=|usr=}}{{25894090}} gab.com Text CRISPR/Cas9-mediated gene editing in human tripronuclear zygotes JO: Protein Cell http://www.kidney.de/pget.php?&tw=1&pmid=25894090 #FOAvip Genome editing tools such as the clustered regularly interspaced short palindromic repeat (CRISPR)-associated system (Cas) have been widely used to modify genes in model systems including animal zygotes and human cells, and hold tremendous promise for both basic research and clinical applications. To date, a serious knowledge gap remains in our understanding of DNA repair mechanisms in human early embryos, and in the efficiency and potential off-target effects of using technologies such as CRISPR/Cas9 in human pre-implantation embryos. In this report, we used tripronuclear (3PN) zygotes to further investigate CRISPR/Cas9-mediated gene editing in human cells. We found that CRISPR/Cas9 could effectively cleave the endogenous beta-globin gene (HBB). However, the efficiency of homologous recombination directed repair (HDR) of HBB was low and the edited embryos were mosaic. Off-target cleavage was also apparent in these 3PN zygotes as revealed by the T7E1 assay and whole-exome sequencing. Furthermore, the endogenous delta-globin gene (HBD), which is homologous to HBB, competed with exogenous donor oligos to act as the repair template, leading to untoward mutations. Our data also indicated that repair of the HBB locus in these embryos occurred preferentially through the non-crossover HDR pathway. Taken together, our work highlights the pressing need to further improve the fidelity and specificity of the CRISPR/Cas9 platform, a prerequisite for any clinical applications of CRSIPR/Cas9-mediated editing. Twit Text FOAVip CRISPR/Cas9-mediated gene editing in human tripronuclear zygotes ????Protein Cell http://www.kidney.de/pget.php?&tw=1&pmid=25894090 #FOAvip Twit Text # Free Paper on # # # # # LINK http://www.kidney.de/pget.php?&tw=1&pmid=25894090 #FOAvip English Wikipedia <ref>{{Cite pmid|25894090}}</ref> CRISPR/Cas9-mediated gene editing in human tripronuclear zygotes #MMPMID25894090 Liang P; Xu Y; Zhang X; Ding C; Huang R; Zhang Z; Lv J; Xie X; Chen Y; Li Y; Sun Y; Bai Y; Songyang Z; Ma W; Zhou C; Huang J Protein Cell 2015[May]; 6 (5): 363-372 PMID25894090show ga Genome editing tools such as the clustered regularly interspaced short palindromic repeat (CRISPR)-associated system (Cas) have been widely used to modify genes in model systems including animal zygotes and human cells, and hold tremendous promise for both basic research and clinical applications. To date, a serious knowledge gap remains in our understanding of DNA repair mechanisms in human early embryos, and in the efficiency and potential off-target effects of using technologies such as CRISPR/Cas9 in human pre-implantation embryos. In this report, we used tripronuclear (3PN) zygotes to further investigate CRISPR/Cas9-mediated gene editing in human cells. We found that CRISPR/Cas9 could effectively cleave the endogenous beta-globin gene (HBB). However, the efficiency of homologous recombination directed repair (HDR) of HBB was low and the edited embryos were mosaic. Off-target cleavage was also apparent in these 3PN zygotes as revealed by the T7E1 assay and whole-exome sequencing. Furthermore, the endogenous delta-globin gene (HBD), which is homologous to HBB, competed with exogenous donor oligos to act as the repair template, leading to untoward mutations. Our data also indicated that repair of the HBB locus in these embryos occurred preferentially through the non-crossover HDR pathway. Taken together, our work highlights the pressing need to further improve the fidelity and specificity of the CRISPR/Cas9 platform, a prerequisite for any clinical applications of CRSIPR/Cas9-mediated editing. |*Blastocyst[MESH] |*CRISPR-Cas Systems[MESH] |*Zygote[MESH] |Hemoglobins, Abnormal/*genetics/*metabolism[MESH]CRISPR/Cas9-mediated gene editing in human tripronuclear zygotes (epmc).pdf DeepDyve Pubget Overpricing