IRES activity validation and Gli-Luciferase reporter assay

XW Xujia Wu
SX Songhua Xiao
MZ Maolei Zhang
LY Lixuan Yang
JZ Jian Zhong
BL Bo Li
FL Fanying Li
XX Xin Xia
XL Xixi Li
HZ Huangkai Zhou
DL Dawei Liu
NH Nunu Huang
XY Xuesong Yang
FX Feizhe Xiao
NZ Nu Zhang
request Request a Protocol
ask Ask a question
Favorite

293T cells were transfected with empty Circ-RLuc-IRES-Reporter vector, EMCV-IRES vector, IRES wildtype, or deletion vectors and incubated for 48 h for analyzing putative IRES activity. For Gli-luciferase reporter assay, indicated cells were seeded in six-well plates and transfected with 8xGliBS-Luc plasmid combined with pRL-TK vector (10:1 ratio) as an internal control. After 48 h, the cells were rinsed with PBS and subjected to dual luciferase assay. A dual luciferase reporter assay system (Promega, Madison, WI, USA) was used based on the manufacturer’s instructions. Firefly luciferase activity was normalized to Renilla luciferase activity for each sample. Data were from three independent assays.

Do you have any questions about this protocol?

Post your question to gather feedback from the community. We will also invite the authors of this article to respond.

post Post a Question
0 Q&A