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. 2016 Sep 30;45(3):e12. doi: 10.1093/nar/gkw883

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© The Author(s) 2016. Published by Oxford University Press on behalf of Nucleic Acids Research.

This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by-nc/4.0/), which permits non-commercial re-use, distribution, and reproduction in any medium, provided the original work is properly cited. For commercial re-use, please contact journals.permissions@oup.com

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Figure 3. — NOP14-AS1 gene modulation using CRISPRn / CRISPRi and antisense LNA GapmeRs. (A) Schematic representation of the MFSD10 / NOP14-AS1 genomic locus depicting the sgRNAs and ASOs used in this study to target NOP14-AS1. (B) Expression analysis of NOP14-AS1 deletion clones in HEK293 cells. RT-qPCR results for MFSD10 and NOP14-AS1 (amplicon #1 and amplicon #2) normalized to cyclophilin A and Wild Type (WT) cells. ND = not detected. Error bars represent SEM (n = 3). * P < 0.05; ** P < 0.01; *** P < 0.001 compared to WT, unpaired two-sided t-test. (C, D) NCI-H460 cells expressing either dCas9 or dCas9-KRAB were transduced with either Control sgRNA or one of the five indicated sgRNAs targeting NOP14-AS1. RT-qPCR results for NOP14-AS1 amplicon #1 (C) and MFSD10 (D) normalized to cyclophilin A and control sgRNA. Error bars represent SEM (n ≥ 4). *P < 0.05; **P < 0.01; ***P < 0.001 compared to dCas9 / dCas9-KRAB + control sgRNA, unpaired two-sided t-test. (E) NCI-H460 cells were transfected with either a control or two independent antisense LNA GapmeRs against NOP14-AS1. RT-qPCR results for NOP14-AS1 amplicon#1, amplicon#2 and MFSD10 normalized to cyclophilin A and control antisense LNA GapmeR. Error bars represent SEM (n = 4). *P < 0.05; **P < 0.01; ***P < 0.001 compared to control GapmeR, unpaired two-sided t-test.