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. 2014 Nov 20;43(3):e17. doi: 10.1093/nar/gku1198

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© The Author(s) 2014. 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/4.0/), which permits unrestricted reuse, distribution, and reproduction in any medium, provided the original work is properly cited.

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Figure 4. — Generation of UCA1 knockout in HCT-116 cells. (A) Colony formation for vector and UCA1 KO for a mixed pool after puromycin selection. (B) Detection of UCA1 expression in UCA1 KO clones by qRT-PCR. (C) Detection of UCA1 target region in all five clones using genomic DNA as a template and primers UCA1-inside-5.3 and UCA1-inside-3.3. (D) Detection of a single nucleotide insertion (in red) in UCA1 KO#2 and #4, by DNA sequencing. The arrow indicates the cleavage site by Cas9. Underlined GGG is PAM sequence. Values in (A) and (D) are means of ± SE (n = 3). **P < 0.01.