Study of the regulatory elements of the Ovalbumin gene promoter using CRISPR technology in chicken cells

Table 2 DNA constructs used in this study

Construct	Features	Application
pdCas9_silencer -gRNA	hU6 promoter- Silencer_gRNA -sgRNA scaffold-CAG promoter-dCas9-polyA	CRISPRi
pdCas9_CAR-gRNA	hU6 promoter- CAR_gRNA -sgRNA scaffold-CAG promoter-dCas9-polyA	CRISPRi
pdCas9-X	CAG promoter-dCas9-polyA	CRISPRi control
pX459_14	hU6 promoter- NRE_gRNA -sgRNA scaffold-CAG promoter-Cas9-T2A- PuroR-bGH polyA	CRISPR Excision
pX459_15	hU6 promoter- SDRE_gRNA -sgRNA scaffold-CAG promoter-Cas9-T2A- PuroR-bGH polyA	CRISPR Excision
pX459_6	hU6 promoter- Ova E2_gRNA -sgRNA scaffold-CAG promoter-Cas9-T2A- PuroR-bGH polyA	CRISPR HDR
pHD_4520	Exon 2 LHA-IRES-DsRed2-HSV TK polyA-CMV promoter –PuroR-IRES2-EGFP-SV40 polyA-Exon 2 RHA	CRISPR HDR

hU6 human U6 promoter, sgRNA single guide RNA, PuroR puromycin N-acetyltransferase resistance gene, bGH polyA bovine growth hormone polyadenylation signal, SV40 polyA SV40 polyadenylation signal, IRES internal ribosome entry site, CAR COUP-adjacent repressor site (− 130 to − 100) in the negative regulatory element (NRE; − 308 to − 88), SDRE steroid-dependent regulatory element (− 900 to − 732), LHA left homology arm, RHA right homology arm

ISSN: 1754-1611