Construction of a combinatorial library of chimeric tumor-specific promoters

Biotechniques. 2017 Sep 1;63(3):107-116. doi: 10.2144/000114586.


Gene therapy is a fast-developing field of molecular medicine. New, effective, and cancer-specific promoters are in high demand by researchers seeking to treat cancer through expression of therapeutic genes. Here, we created a combinatorial library of tumor-specific chimeric promoter modules for identifying new promoters with desired functions. The library was constructed by randomly combining promoter fragments from eight human genes involved in cell proliferation control. The pool of chimeric promoters was inserted into a lentiviral expression vector upstream of the CopGFP reporter gene, transduced into A431 cells, and enriched for active promoters by cell sorting. The enriched library contained a remarkably high proportion of active and tumor-specific promoters. This approach to generating combinatorial libraries of chimeric promoters may serve as a useful tool for selecting highly specific and effective promoters for cancer research and gene therapy.

Keywords: artificial promoter; cancer gene therapy; cancer specific; chimeric promoter; random promoter library.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Cell Line, Tumor
  • Cell Proliferation / genetics*
  • Fibroblasts / cytology
  • Gene Library*
  • Genetic Therapy / methods*
  • Genetic Vectors
  • HEK293 Cells
  • Humans
  • Lentivirus / genetics
  • Neoplasms / genetics*
  • Neoplasms / therapy*
  • Primary Cell Culture
  • Promoter Regions, Genetic / genetics*
  • Transfection