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10.1038/nmeth.3360 http://scihub22266oqcxt.onion/10.1038/nmeth.3360 C4667794!4667794!25867848     free     free     free Warning: file_get_contents(https://eutils.ncbi.nlm.nih.gov/entrez/eutils/elink.fcgi?dbfrom=pubmed&id=25867848&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 209.6 to int loses precision in C:\Inetpub\vhosts\kidney.de\httpdocs\pget.php on line 534       Nat+Methods 2015 ; 12 (6): 535-40 Nephropedia Template TP {{tp|p=25867848|t=2015. Rapid reverse genetic screening using CRISPR in zebrafish |pdf=|usr=}}{{25867848}} gab.com Text Rapid reverse genetic screening using CRISPR in zebrafish JO: Nat Methods http://www.kidney.de/pget.php?&tw=1&pmid=25867848 #FOAvip Identifying genes involved in biological processes is critical for understanding the molecular building blocks of life. The effectiveness of engineered CRISPR (Clustered Regularly Interspaced Short Palindromic Repeats) to efficiently mutate specific loci coupled with the accessibility of zebrafish (Danio rerio) provides an opportunity to screen for genes involved in vertebrate biological processes. Injection of Cas9-encoding mRNA and an engineered, single guide RNA (sgRNA) can cause biallelic mutations in injected embryos that phenocopy known mutant phenotypes. We found that increasing CRISPR efficiency and multiplexing sgRNAs allowed for phenocopy of known mutants across many phenotypes. We performed a proof-of-concept screen examining 48 loci by intersecting, multiplexed pool injections, and identified two new genes involved in electrical synapse formation. By deep-sequencing target loci we found that 90% of the genes were effectively screened. We conclude that CRISPR can be used as a powerful reverse genetic screening strategy in vivo in a vertebrate system. Twit Text FOAVip Rapid reverse genetic screening using CRISPR in zebrafish ????Nat Methods http://www.kidney.de/pget.php?&tw=1&pmid=25867848 #FOAvip Twit Text # Free Paper on # # # # # LINK http://www.kidney.de/pget.php?&tw=1&pmid=25867848 #FOAvip English Wikipedia <ref>{{Cite pmid|25867848}}</ref> Rapid reverse genetic screening using CRISPR in zebrafish #MMPMID25867848 Shah AN; Davey CF; Whitebirch AC; Miller AC; Moens CB Nat Methods 2015[Jun]; 12 (6): 535-40 PMID25867848show ga Identifying genes involved in biological processes is critical for understanding the molecular building blocks of life. The effectiveness of engineered CRISPR (Clustered Regularly Interspaced Short Palindromic Repeats) to efficiently mutate specific loci coupled with the accessibility of zebrafish (Danio rerio) provides an opportunity to screen for genes involved in vertebrate biological processes. Injection of Cas9-encoding mRNA and an engineered, single guide RNA (sgRNA) can cause biallelic mutations in injected embryos that phenocopy known mutant phenotypes. We found that increasing CRISPR efficiency and multiplexing sgRNAs allowed for phenocopy of known mutants across many phenotypes. We performed a proof-of-concept screen examining 48 loci by intersecting, multiplexed pool injections, and identified two new genes involved in electrical synapse formation. By deep-sequencing target loci we found that 90% of the genes were effectively screened. We conclude that CRISPR can be used as a powerful reverse genetic screening strategy in vivo in a vertebrate system. äRapid reverse genetic screening using CRISPR in zebrafish (epmc).pdf DeepDyve Pubget Overpricing