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10.1091/mbc.E17-05-0288 http://scihub22266oqcxt.onion/10.1091/mbc.E17-05-0288 C5909934!5909934 !29167381     free     free     free Warning: file_get_contents(https://eutils.ncbi.nlm.nih.gov/entrez/eutils/elink.fcgi?dbfrom=pubmed&id=29167381 &cmd=llinks): Failed to open stream: HTTP request failed! 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CRISPR-Trap: a clean approach for the generation of gene knockouts and gene replacements in human cells |pdf=|usr=}}{{29167381 }} gab.com Text CRISPR-Trap: a clean approach for the generation of gene knockouts and gene replacements in human cells JO: Mol Biol Cell http://www.kidney.de/pget.php?&tw=1&pmid=29167381 #FOAvip CRISPR/Cas9-based genome editing offers the possibility to knock out almost any gene of interest in an affordable and simple manner. The most common strategy is the introduction of a frameshift into the open reading frame (ORF) of the target gene which truncates the coding sequence (CDS) and targets the corresponding transcript for degradation by nonsense-mediated mRNA decay (NMD). However, we show that transcripts containing premature termination codons (PTCs) are not always degraded efficiently and can generate C-terminally truncated proteins which might have residual or dominant negative functions. Therefore, we recommend an alternative approach for knocking out genes, which combines CRISPR/Cas9 with gene traps (CRISPR-Trap) and is applicable to ?50% of all spliced human protein-coding genes and a large subset of lncRNAs. CRISPR-Trap completely prevents the expression of the ORF and avoids expression of C-terminal truncated proteins. We demonstrate the feasibility of CRISPR-Trap by utilizing it to knock out several genes in different human cell lines. Finally, we also show that this approach can be used to efficiently generate gene replacements allowing for modulation of protein levels for otherwise lethal knockouts (KOs). Thus, CRISPR-Trap offers several advantages over conventional KO approaches and allows for generation of clean CRISPR/Cas9-based KOs. Twit Text FOAVip CRISPR-Trap: a clean approach for the generation of gene knockouts and gene replacements in human cells ????Mol Biol Cell http://www.kidney.de/pget.php?&tw=1&pmid=29167381 #FOAvip Twit Text # Free Paper on # # # # # LINK http://www.kidney.de/pget.php?&tw=1&pmid=29167381 #FOAvip English Wikipedia <ref>{{Cite pmid|29167381 }}</ref> CRISPR-Trap: a clean approach for the generation of gene knockouts and gene replacements in human cells #MMPMID29167381 Reber S ; Mechtersheimer J ; Nasif S ; Benitez JA ; Colombo M ; Domanski M ; Jutzi D ; Hedlund E ; Ruepp MD Mol Biol Cell 2018[Jan]; 29 (2 ): 75-83 PMID29167381 show ga CRISPR/Cas9-based genome editing offers the possibility to knock out almost any gene of interest in an affordable and simple manner. The most common strategy is the introduction of a frameshift into the open reading frame (ORF) of the target gene which truncates the coding sequence (CDS) and targets the corresponding transcript for degradation by nonsense-mediated mRNA decay (NMD). However, we show that transcripts containing premature termination codons (PTCs) are not always degraded efficiently and can generate C-terminally truncated proteins which might have residual or dominant negative functions. Therefore, we recommend an alternative approach for knocking out genes, which combines CRISPR/Cas9 with gene traps (CRISPR-Trap) and is applicable to ?50% of all spliced human protein-coding genes and a large subset of lncRNAs. CRISPR-Trap completely prevents the expression of the ORF and avoids expression of C-terminal truncated proteins. We demonstrate the feasibility of CRISPR-Trap by utilizing it to knock out several genes in different human cell lines. Finally, we also show that this approach can be used to efficiently generate gene replacements allowing for modulation of protein levels for otherwise lethal knockouts (KOs). Thus, CRISPR-Trap offers several advantages over conventional KO approaches and allows for generation of clean CRISPR/Cas9-based KOs. |*CRISPR-Cas Systems [MESH] |*Clustered Regularly Interspaced Short Palindromic Repeats [MESH] |Gene Knockout Techniques/*methods [MESH] |HEK293 Cells [MESH] |HeLa Cells [MESH]CRISPR-Trap: a clean approach for the generation of gene knockouts and(epmc).pdf DeepDyve Pubget Overpricing