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10.1038/srep32386 http://scihub22266oqcxt.onion/10.1038/srep32386 C5004112!5004112 !27572667     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 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 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\27572667 .jpg): Failed to open stream: No such file or directory in C:\Inetpub\vhosts\kidney.de\httpdocs\pget.php on line 117       Sci+Rep 2016 ; 6 (ä): 32386 Nephropedia Template TP {{tp|p=27572667 |t=2016. In vivo mutagenesis of miRNA gene families using a scalable multiplexed CRISPR/Cas9 nuclease system |pdf=|usr=}}{{27572667 }} gab.com Text In vivo mutagenesis of miRNA gene families using a scalable multiplexed CRISPR/Cas9 nuclease system JO: Sci Rep http://www.kidney.de/pget.php?&tw=1&pmid=27572667 #FOAvip A large number of microRNAs (miRNAs) are grouped into families derived from the same phylogenetic ancestors. miRNAs within a family often share the same physiological functions despite differences in their primary sequences, secondary structures, or chromosomal locations. Consequently, the generation of animal models to analyze the activity of miRNA families is extremely challenging. Using zebrafish as a model system, we successfully provide experimental evidence that a large number of miRNAs can be simultaneously mutated to abrogate the activity of an entire miRNA family. We show that injection of the Cas9 nuclease and two, four, ten, and up to twenty-four multiplexed single guide RNAs (sgRNAs) can induce mutations in 90% of the miRNA genomic sequences analyzed. We performed a survey of these 45 mutations in 10 miRNA genes, analyzing the impact of our mutagenesis strategy on the processing of each miRNA both computationally and in vivo. Our results offer an effective approach to mutate and study the activity of miRNA families and pave the way for further analysis on the function of complex miRNA families in higher multicellular organisms. Twit Text FOAVip In vivo mutagenesis of miRNA gene families using a scalable multiplexed CRISPR/Cas9 nuclease system ????Sci Rep http://www.kidney.de/pget.php?&tw=1&pmid=27572667 #FOAvip Twit Text # Free Paper on # # # # # LINK http://www.kidney.de/pget.php?&tw=1&pmid=27572667 #FOAvip English Wikipedia <ref>{{Cite pmid|27572667 }}</ref> In vivo mutagenesis of miRNA gene families using a scalable multiplexed CRISPR/Cas9 nuclease system #MMPMID27572667 Narayanan A ; Hill-Teran G ; Moro A ; Ristori E ; Kasper DM ; A Roden C ; Lu J ; Nicoli S Sci Rep 2016[Aug]; 6 (ä): 32386 PMID27572667 show ga A large number of microRNAs (miRNAs) are grouped into families derived from the same phylogenetic ancestors. miRNAs within a family often share the same physiological functions despite differences in their primary sequences, secondary structures, or chromosomal locations. Consequently, the generation of animal models to analyze the activity of miRNA families is extremely challenging. Using zebrafish as a model system, we successfully provide experimental evidence that a large number of miRNAs can be simultaneously mutated to abrogate the activity of an entire miRNA family. We show that injection of the Cas9 nuclease and two, four, ten, and up to twenty-four multiplexed single guide RNAs (sgRNAs) can induce mutations in 90% of the miRNA genomic sequences analyzed. We performed a survey of these 45 mutations in 10 miRNA genes, analyzing the impact of our mutagenesis strategy on the processing of each miRNA both computationally and in vivo. Our results offer an effective approach to mutate and study the activity of miRNA families and pave the way for further analysis on the function of complex miRNA families in higher multicellular organisms. |Animals [MESH] |CRISPR-Cas Systems/*genetics [MESH] |Chromosomes/genetics [MESH] |Genome/genetics [MESH] |MicroRNAs/*genetics [MESH] |Multigene Family/*genetics [MESH] |Mutagenesis/*genetics [MESH] |Mutation [MESH]In vivo mutagenesis of miRNA gene families using a scalable multiplexe(epmc).pdf DeepDyve Pubget Overpricing