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10.1101/gr.231936.117 http://scihub22266oqcxt.onion/10.1101/gr.231936.117 C5848615!5848615!29472270     free     free     free Warning: file_get_contents(https://eutils.ncbi.nlm.nih.gov/entrez/eutils/elink.fcgi?dbfrom=pubmed&id=29472270&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 227.6 to int loses precision in C:\Inetpub\vhosts\kidney.de\httpdocs\pget.php on line 534 Deprecated: Implicit conversion from float 227.6 to int loses precision in C:\Inetpub\vhosts\kidney.de\httpdocs\pget.php on line 534       Genome+Res 2018 ; 28 (3): 367-73 Nephropedia Template TP {{tp|p=29472270|t=2018. CRISPR RNAs trigger innate immune responses in human cells |pdf=|usr=}}{{29472270}} gab.com Text CRISPR RNAs trigger innate immune responses in human cells JO: Genome Res http://www.kidney.de/pget.php?&tw=1&pmid=29472270 #FOAvip Here, we report that CRISPR guide RNAs (gRNAs) with a 5?-triphosphate group (5?-ppp gRNAs) produced via in vitro transcription trigger RNA-sensing innate immune responses in human and murine cells, leading to cytotoxicity. 5?-ppp gRNAs in the cytosol are recognized by DDX58, which in turn activates type I interferon responses, causing up to ?80% cell death. We show that the triphosphate group can be removed by a phosphatase in vitro and that the resulting 5?-hydroxyl gRNAs in complex with Cas9 or Cpf1 avoid innate immune responses and can achieve targeted mutagenesis at a frequency of 95% in primary human CD4+ T cells. These results are in line with previous findings that chemically synthesized sgRNAs with a 5?-hydroxyl group are much more efficient than in vitro?transcribed (IVT) sgRNAs in human and other mammalian cells. The phosphatase treatment of IVT sgRNAs is a cost-effective method for making highly active sgRNAs, avoiding innate immune responses in human cells. Twit Text FOAVip CRISPR RNAs trigger innate immune responses in human cells ????Genome Res http://www.kidney.de/pget.php?&tw=1&pmid=29472270 #FOAvip Twit Text # Free Paper on # # # # # LINK http://www.kidney.de/pget.php?&tw=1&pmid=29472270 #FOAvip English Wikipedia <ref>{{Cite pmid|29472270}}</ref> CRISPR RNAs trigger innate immune responses in human cells #MMPMID29472270 Kim S; Koo T; Jee HG; Cho HY; Lee G; Lim DG; Shin HS; Kim JS Genome Res 2018[Mar]; 28 (3): 367-73 PMID29472270show ga Here, we report that CRISPR guide RNAs (gRNAs) with a 5?-triphosphate group (5?-ppp gRNAs) produced via in vitro transcription trigger RNA-sensing innate immune responses in human and murine cells, leading to cytotoxicity. 5?-ppp gRNAs in the cytosol are recognized by DDX58, which in turn activates type I interferon responses, causing up to ?80% cell death. We show that the triphosphate group can be removed by a phosphatase in vitro and that the resulting 5?-hydroxyl gRNAs in complex with Cas9 or Cpf1 avoid innate immune responses and can achieve targeted mutagenesis at a frequency of 95% in primary human CD4+ T cells. These results are in line with previous findings that chemically synthesized sgRNAs with a 5?-hydroxyl group are much more efficient than in vitro?transcribed (IVT) sgRNAs in human and other mammalian cells. The phosphatase treatment of IVT sgRNAs is a cost-effective method for making highly active sgRNAs, avoiding innate immune responses in human cells. äCRISPR RNAs trigger innate immune responses in human cells (epmc).pdf DeepDyve Pubget Overpricing