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10.1038/s41467-018-03044-y http://scihub22266oqcxt.onion/10.1038/s41467-018-03044-y C5838097!5838097 !29507284     free     free     free Warning: file_get_contents(https://eutils.ncbi.nlm.nih.gov/entrez/eutils/elink.fcgi?dbfrom=pubmed&id=29507284 &cmd=llinks): Failed to open stream: HTTP request failed! HTTP/1.1 429 Too Many Requests in C:\Inetpub\vhosts\kidney.de\httpdocs\pget.php on line 215 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 Warning: imagejpeg(C:\Inetpub\vhosts\kidney.de\httpdocs\phplern\29507284 .jpg): Failed to open stream: No such file or directory in C:\Inetpub\vhosts\kidney.de\httpdocs\pget.php on line 117       Nat+Commun 2018 ; 9 (1 ): 939 Nephropedia Template TP {{tp|p=29507284 |t=2018. Microhomology-assisted scarless genome editing in human iPSCs |pdf=|usr=}}{{29507284 }} gab.com Text Microhomology-assisted scarless genome editing in human iPSCs JO: Nat Commun http://www.kidney.de/pget.php?&tw=1&pmid=29507284 #FOAvip Gene-edited induced pluripotent stem cells (iPSCs) provide relevant isogenic human disease models in patient-specific or healthy genetic backgrounds. Towards this end, gene targeting using antibiotic selection along with engineered point mutations remains a reliable method to enrich edited cells. Nevertheless, integrated selection markers obstruct scarless transgene-free gene editing. Here, we present a method for scarless selection marker excision using engineered microhomology-mediated end joining (MMEJ). By overlapping the homology arms of standard donor vectors, short tandem microhomologies are generated flanking the selection marker. Unique CRISPR-Cas9 protospacer sequences nested between the selection marker and engineered microhomologies are cleaved after gene targeting, engaging MMEJ and scarless excision. Moreover, when point mutations are positioned unilaterally within engineered microhomologies, both mutant and normal isogenic clones are derived simultaneously. The utility and fidelity of our method is demonstrated in human iPSCs by editing the X-linked HPRT1 locus and biallelic modification of the autosomal APRT locus, eliciting disease-relevant metabolic phenotypes. Twit Text FOAVip Microhomology-assisted scarless genome editing in human iPSCs ????Nat Commun http://www.kidney.de/pget.php?&tw=1&pmid=29507284 #FOAvip Twit Text # Free Paper on # # # # # LINK http://www.kidney.de/pget.php?&tw=1&pmid=29507284 #FOAvip English Wikipedia <ref>{{Cite pmid|29507284 }}</ref> Microhomology-assisted scarless genome editing in human iPSCs #MMPMID29507284 Kim SI ; Matsumoto T ; Kagawa H ; Nakamura M ; Hirohata R ; Ueno A ; Ohishi M ; Sakuma T ; Soga T ; Yamamoto T ; Woltjen K Nat Commun 2018[Mar]; 9 (1 ): 939 PMID29507284 show ga Gene-edited induced pluripotent stem cells (iPSCs) provide relevant isogenic human disease models in patient-specific or healthy genetic backgrounds. Towards this end, gene targeting using antibiotic selection along with engineered point mutations remains a reliable method to enrich edited cells. Nevertheless, integrated selection markers obstruct scarless transgene-free gene editing. Here, we present a method for scarless selection marker excision using engineered microhomology-mediated end joining (MMEJ). By overlapping the homology arms of standard donor vectors, short tandem microhomologies are generated flanking the selection marker. Unique CRISPR-Cas9 protospacer sequences nested between the selection marker and engineered microhomologies are cleaved after gene targeting, engaging MMEJ and scarless excision. Moreover, when point mutations are positioned unilaterally within engineered microhomologies, both mutant and normal isogenic clones are derived simultaneously. The utility and fidelity of our method is demonstrated in human iPSCs by editing the X-linked HPRT1 locus and biallelic modification of the autosomal APRT locus, eliciting disease-relevant metabolic phenotypes. |*DNA End-Joining Repair [MESH] |*Gene Editing [MESH] |Alleles [MESH] |Amino Acid Sequence [MESH] |Base Sequence [MESH] |Chromosomes, Human/genetics [MESH] |Genetic Loci [MESH] |HEK293 Cells [MESH] |Humans [MESH] |Induced Pluripotent Stem Cells/*metabolism [MESH] |Mutation/genetics [MESH]Microhomology-assisted scarless genome editing in human iPSCs (epmc).pdf DeepDyve Pubget Overpricing