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10.1242/bio.201410009 http://scihub22266oqcxt.onion/10.1242/bio.201410009 C4365486!4365486 !25596277     free     free     free Warning: file_get_contents(https://eutils.ncbi.nlm.nih.gov/entrez/eutils/elink.fcgi?dbfrom=pubmed&id=25596277 &cmd=llinks): Failed to open stream: HTTP request failed! 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Highly efficient gene knockout by injection of TALEN mRNAs into oocytes and host transfer in Xenopus laevis |pdf=|usr=}}{{25596277 }} gab.com Text Highly efficient gene knockout by injection of TALEN mRNAs into oocytes and host transfer in Xenopus laevis JO: Biol Open http://www.kidney.de/pget.php?&tw=1&pmid=25596277 #FOAvip Zinc-finger nucleases, transcription activator-like effector nucleases (TALENs) and the CRISPR/Cas (clustered regularly interspaced short palindromic repeats/CRISPR-associated proteins) system are potentially powerful tools for producing tailor-made knockout animals. However, their mutagenic activity is not high enough to induce mutations at all loci of a target gene throughout an entire tadpole. In this study, we present a highly efficient method for introducing gene modifications at almost all target sequences in randomly selected embryos. The gene modification activity of TALEN is enhanced by adopting the host-transfer technique. In our method, the efficiency is further improved by injecting TALEN mRNAs fused to the 3'UTR of the Xenopus DEADSouth gene into oocytes, which are then transferred into a host female frog, where they are ovulated and fertilized. The addition of the 3'UTR of the DEADSouth gene promotes mRNA translation in the oocytes and increases the expression of TALEN proteins to near-maximal levels three hours post fertilization (hpf). In contrast, TALEN mRNAs without this 3'UTR are translated infrequently in oocytes. Our data suggest that genomic DNA is more sensitive to TALEN proteins from fertilization to the midblastula (MBT) stage. Our method works by increasing the levels of TALEN proteins during the pre-MBT stages. Twit Text FOAVip Highly efficient gene knockout by injection of TALEN mRNAs into oocytes and host transfer in Xenopus laevis ????Biol Open http://www.kidney.de/pget.php?&tw=1&pmid=25596277 #FOAvip Twit Text # Free Paper on # # # # # LINK http://www.kidney.de/pget.php?&tw=1&pmid=25596277 #FOAvip English Wikipedia <ref>{{Cite pmid|25596277 }}</ref> Highly efficient gene knockout by injection of TALEN mRNAs into oocytes and host transfer in Xenopus laevis #MMPMID25596277 Nakajima K ; Yaoita Y Biol Open 2015[Jan]; 4 (2 ): 180-5 PMID25596277 show ga Zinc-finger nucleases, transcription activator-like effector nucleases (TALENs) and the CRISPR/Cas (clustered regularly interspaced short palindromic repeats/CRISPR-associated proteins) system are potentially powerful tools for producing tailor-made knockout animals. However, their mutagenic activity is not high enough to induce mutations at all loci of a target gene throughout an entire tadpole. In this study, we present a highly efficient method for introducing gene modifications at almost all target sequences in randomly selected embryos. The gene modification activity of TALEN is enhanced by adopting the host-transfer technique. In our method, the efficiency is further improved by injecting TALEN mRNAs fused to the 3'UTR of the Xenopus DEADSouth gene into oocytes, which are then transferred into a host female frog, where they are ovulated and fertilized. The addition of the 3'UTR of the DEADSouth gene promotes mRNA translation in the oocytes and increases the expression of TALEN proteins to near-maximal levels three hours post fertilization (hpf). In contrast, TALEN mRNAs without this 3'UTR are translated infrequently in oocytes. Our data suggest that genomic DNA is more sensitive to TALEN proteins from fertilization to the midblastula (MBT) stage. Our method works by increasing the levels of TALEN proteins during the pre-MBT stages. äHighly efficient gene knockout by injection of TALEN mRNAs into oocyte(epmc).pdf DeepDyve Pubget Overpricing