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10.1186/s12864-015-2086-z http://scihub22266oqcxt.onion/10.1186/s12864-015-2086-z C4619085!4619085!26493208     free     free     free Deprecated: Implicit conversion from float 215.6 to int loses precision in C:\Inetpub\vhosts\kidney.de\httpdocs\pget.php on line 534 Deprecated: Implicit conversion from float 215.6 to int loses precision in C:\Inetpub\vhosts\kidney.de\httpdocs\pget.php on line 534 Deprecated: Implicit conversion from float 215.6 to int loses precision in C:\Inetpub\vhosts\kidney.de\httpdocs\pget.php on line 534 Deprecated: Implicit conversion from float 215.6 to int loses precision in C:\Inetpub\vhosts\kidney.de\httpdocs\pget.php on line 534 Deprecated: Implicit conversion from float 215.6 to int loses precision in C:\Inetpub\vhosts\kidney.de\httpdocs\pget.php on line 534       BMC+Genomics 2015 ; 16 (ä): ä Nephropedia Template TP {{tp|p=26493208|t=2015. DECKO: Single-oligo, dual-CRISPR deletion of genomic elements including long non-coding RNAs |pdf=|usr=}}{{26493208}} gab.com Text DECKO: Single-oligo, dual-CRISPR deletion of genomic elements including long non-coding RNAs JO: BMC Genomics http://www.kidney.de/pget.php?&tw=1&pmid=26493208 #FOAvip Background: CRISPR genome-editing technology makes it possible to quickly and cheaply delete non-protein-coding regulatory elements. We present a vector system adapted for this purpose called DECKO (Double Excision CRISPR Knockout), which applies a simple two-step cloning to generate lentiviral vectors expressing two guide RNAs (gRNAs) simultaneously. The key feature of DECKO is its use of a single 165 bp starting oligonucleotide carrying the variable sequences of both gRNAs, making it fully scalable from single-locus studies to complex library cloning. Results: We apply DECKO to deleting the promoters of one protein-coding gene and two oncogenic lncRNAs, UCA1 and the highly-expressed MALAT1, focus of many previous studies employing RNA interference approaches. DECKO successfully deleted genomic fragments ranging in size from 100 to 3000 bp in four human cell lines. Using a clone-derivation workflow lasting approximately 20 days, we obtained 9 homozygous and 17 heterozygous promoter knockouts in three human cell lines. Frequent target region inversions were observed. These clones have reductions in steady-state MALAT1 RNA levels of up to 98 % and display reduced proliferation rates. Conclusions: We present a dual CRISPR tool, DECKO, which is cloned using a single starting oligonucleotide, thereby affording simplicity and scalability to CRISPR knockout studies of non-coding genomic elements, including long non-coding RNAs. Electronic supplementary material: The online version of this article (doi:10.1186/s12864-015-2086-z) contains supplementary material, which is available to authorized users. Twit Text FOAVip DECKO: Single-oligo, dual-CRISPR deletion of genomic elements including long non-coding RNAs ????BMC Genomics http://www.kidney.de/pget.php?&tw=1&pmid=26493208 #FOAvip Twit Text # Free Paper on # # # # # LINK http://www.kidney.de/pget.php?&tw=1&pmid=26493208 #FOAvip English Wikipedia <ref>{{Cite pmid|26493208}}</ref> DECKO: Single-oligo, dual-CRISPR deletion of genomic elements including long non-coding RNAs #MMPMID26493208 Aparicio-Prat E; Arnan C; Sala I; Bosch N; Guigó R; Johnson R BMC Genomics 2015[]; 16 (ä): ä PMID26493208show ga Background: CRISPR genome-editing technology makes it possible to quickly and cheaply delete non-protein-coding regulatory elements. We present a vector system adapted for this purpose called DECKO (Double Excision CRISPR Knockout), which applies a simple two-step cloning to generate lentiviral vectors expressing two guide RNAs (gRNAs) simultaneously. The key feature of DECKO is its use of a single 165 bp starting oligonucleotide carrying the variable sequences of both gRNAs, making it fully scalable from single-locus studies to complex library cloning. Results: We apply DECKO to deleting the promoters of one protein-coding gene and two oncogenic lncRNAs, UCA1 and the highly-expressed MALAT1, focus of many previous studies employing RNA interference approaches. DECKO successfully deleted genomic fragments ranging in size from 100 to 3000 bp in four human cell lines. Using a clone-derivation workflow lasting approximately 20 days, we obtained 9 homozygous and 17 heterozygous promoter knockouts in three human cell lines. Frequent target region inversions were observed. These clones have reductions in steady-state MALAT1 RNA levels of up to 98 % and display reduced proliferation rates. Conclusions: We present a dual CRISPR tool, DECKO, which is cloned using a single starting oligonucleotide, thereby affording simplicity and scalability to CRISPR knockout studies of non-coding genomic elements, including long non-coding RNAs. Electronic supplementary material: The online version of this article (doi:10.1186/s12864-015-2086-z) contains supplementary material, which is available to authorized users. äDECKO: Single-oligo, dual-CRISPR deletion of genomic elements includin(epmc).pdf DeepDyve Pubget Overpricing