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10.1016/j.ymeth.2014.05.003 http://scihub22266oqcxt.onion/10.1016/j.ymeth.2014.05.003 C4405113!4405113!24929070     free     free     free Deprecated: Implicit conversion from float 211.6 to int loses precision in C:\Inetpub\vhosts\kidney.de\httpdocs\pget.php on line 534 Deprecated: Implicit conversion from float 211.6 to int loses precision in C:\Inetpub\vhosts\kidney.de\httpdocs\pget.php on line 534 Deprecated: Implicit conversion from float 211.6 to int loses precision in C:\Inetpub\vhosts\kidney.de\httpdocs\pget.php on line 534 Deprecated: Implicit conversion from float 211.6 to int loses precision in C:\Inetpub\vhosts\kidney.de\httpdocs\pget.php on line 534 Deprecated: Implicit conversion from float 211.6 to int loses precision in C:\Inetpub\vhosts\kidney.de\httpdocs\pget.php on line 534 Deprecated: Implicit conversion from float 211.6 to int loses precision in C:\Inetpub\vhosts\kidney.de\httpdocs\pget.php on line 534 Deprecated: Implicit conversion from float 211.6 to int loses precision in C:\Inetpub\vhosts\kidney.de\httpdocs\pget.php on line 534       Methods 2014 ; 69 (2): 171-8 Nephropedia Template TP {{tp|p=24929070|t=2014. Biallelic targeting of expressed genes in mouse embryonic stem cells using the Cas9 system |pdf=|usr=}}{{24929070}} gab.com Text Biallelic targeting of expressed genes in mouse embryonic stem cells using the Cas9 system JO: Methods http://www.kidney.de/pget.php?&tw=1&pmid=24929070 #FOAvip Gene targeting ? homologous recombination between transfected DNA and a chromosomal locus ? is greatly stimulated by a DNA break in the target locus. Recently, the RNA-guided Cas9 endonuclease, involved in bacterial adaptive immunity, has been modified to function in mammalian cells. Unlike other site-specific endonucleases whose specificity resides within a protein, the specificity of Cas9-mediated DNA cleavage is determined by a guide RNA (gRNA) containing an ~20 nucleotide locus-specific RNA sequence, representing a major advance for versatile site-specific cleavage of the genome without protein engineering. This article provides a detailed method using the Cas9 system to target expressed genes in mouse embryonic stem cells. In this method, a promoterless marker flanked by short homology arms to the target locus is transfected into cells together with Cas9 and gRNA expression vectors. Importantly, biallelic gene knockout is obtained at high frequency by only one round of targeting using a single marker. Twit Text FOAVip Biallelic targeting of expressed genes in mouse embryonic stem cells using the Cas9 system ????Methods http://www.kidney.de/pget.php?&tw=1&pmid=24929070 #FOAvip Twit Text # Free Paper on # # # # # LINK http://www.kidney.de/pget.php?&tw=1&pmid=24929070 #FOAvip English Wikipedia <ref>{{Cite pmid|24929070}}</ref> Biallelic targeting of expressed genes in mouse embryonic stem cells using the Cas9 system #MMPMID24929070 Zhang Y; Vanoli F; LaRocque JR; Krawczyk PM; Jasin M Methods 2014[Sep]; 69 (2): 171-8 PMID24929070show ga Gene targeting ? homologous recombination between transfected DNA and a chromosomal locus ? is greatly stimulated by a DNA break in the target locus. Recently, the RNA-guided Cas9 endonuclease, involved in bacterial adaptive immunity, has been modified to function in mammalian cells. Unlike other site-specific endonucleases whose specificity resides within a protein, the specificity of Cas9-mediated DNA cleavage is determined by a guide RNA (gRNA) containing an ~20 nucleotide locus-specific RNA sequence, representing a major advance for versatile site-specific cleavage of the genome without protein engineering. This article provides a detailed method using the Cas9 system to target expressed genes in mouse embryonic stem cells. In this method, a promoterless marker flanked by short homology arms to the target locus is transfected into cells together with Cas9 and gRNA expression vectors. Importantly, biallelic gene knockout is obtained at high frequency by only one round of targeting using a single marker. äBiallelic targeting of expressed genes in mouse embryonic stem cells u(epmc).pdf DeepDyve Pubget Overpricing