1. Academic Validation
  2. Combining protein and RNA quantification to evaluate promoter activity by using dual-color fluorescent reporting systems

Combining protein and RNA quantification to evaluate promoter activity by using dual-color fluorescent reporting systems

  • Biosci Rep. 2021 Sep 30;41(9):BSR20211525. doi: 10.1042/BSR20211525.
Yan Peng 1 2 Xin Huang 1 Tianfang Huang 1 Feng Du 1 Xin Cui 1 Zhuo Tang 1
Affiliations

Affiliations

  • 1 Natural Products Research Center, Chengdu Institute of Biology, Chinese Academy of Sciences, Chengdu, Sichuan 610041, P.R. China.
  • 2 Savaid Medical School, University of Chinese Academy of Sciences, Beijing 100049, P.R. China.
Abstract

Herein, Broccoli/mCherry and an EGFP/mCherry dual-color fluorescent reporting systems have been established to quantify the promoter activity at transcription and translation levels in eukaryotic cells. Based on those systems, four commonly used promoters (CMV and SV40 of Pol II and U6, H1 of Pol III) were accurately evaluated at both the transcriptional and translational levels by combining accurate protein and RNA quantification. Furthermore, we verified that Pol III promoters can induce proteins expression, and Pol II promoter can be applied to express RNA molecules with defined length by combining a self-cleaving ribozyme and an artificial poly(A) tail. The dual-color fluorescence reporting systems described here could play a significant role in evaluating other gene expression regulators for gene therapy.

Keywords

Broccoli; Promoter; dual-color; fluorescent protein; transcription; translation.

Figures
Products
  • Cat. No.
    Product Name
    Description
    Target
    Research Area
  • HY-110251
    98.12%, RNA Aptamers Probe