Table 5.

DNA constructs used in this study

Construct	Features	Application
pX459_2	hU6 promoter- sgRNA (downstream of CCAT1 Exon 1)-sgRNA scaffold-CAG promoter-Cas9-T2A- PuroR-bGH polyA	CRISPR Excision CRISPR du-HITI
pX459_3	hU6 promoter- sgRNA (upstream of CCAT1 Exon 1)-sgRNA scaffold-CAG promoter-Cas9-T2A- PuroR-bGH polyA	CRISPR Excision CRISPR du-HITI
pX459_13	hU6 promoter- sgRNA (CCAT1 Exon 2)-sgRNA scaffold-CAG promoter-Cas9-T2A- PuroR-bGH polyA	CRISPR HDR
pHD_4317_CCAT1 E2 HAs	Exon 2 LHA-DsRed2-HSV TK polyA-CMV promoter –PuroR-IRES2-EGFP-SV40 polyA-Exon 2 RHA	CRISPR HDR
pX460_11	hU6 promoter-sgRNA (downstream of CCAT1 Exon 1) plus PAM-sgRNA scaffold-CAG promoter-EGFP-bGH polyA	CRISPR du-HITI
pX461_11	hU6 promoter-sgRNA (downstream of CCAT1 Exon 1) plus PAM-sgRNA scaffold-CAG promoter-PuroR-bGH polyA	CRISPR du-HITI

hU6 human U6 promoter, sgRNA single guide RNA, PuroR puromycin N-acetyltransferase, bGH bovine growth hormone polyadenylation signal, SV40 polyA SV40 polyadenylation signal, IRES internal ribosome entry site