Conversion of a Non-Cancer-Selective Promoter into a Cancer-Selective Promoter
Abstract
Progression-elevated gene-3 (PEG-3) and rat growth arrest and DNA damage-inducible gene-34 (GADD34) display significant sequence homology with regulation predominantly transcriptional. The rat full-length (FL) and minimal (min) PEG-3 promoter display cancer-selective expression in rodent and human tumors, allowing for cancer-directed regulation of transgenes, viral replication and in vivo imaging of tumors and metastases in animals, whereas the FL- and min-GADD34-Prom lack cancer specificity. Min-PEG-Prom and min-GADD34-Prom have identical sequences except for two single-point mutation differences (at −260 bp and +159 bp). Engineering double mutations in the min-GADD34-Prom produce the GAPE-Prom. Changing one base pair (+159) or both point mutations in the min-GADD34-Prom, but not the FL-GADD34-Prom, results in cancer-selective transgene expression in diverse cancer cells (including prostate, breast, pancreatic and neuroblastoma) vs. normal counterparts. Additionally, we identified a GATA2 transcription factor binding site, promoting cancer specificity when both min-PEG-Prom mutations are present in the GAPE-Prom. Taken together, introducing specific point mutations in a rat min-GADD34-Prom converts this non-cancer-specific promoter into a cancer-selective promoter, and the addition of GATA2 with existing AP1 and PEA3 transcription factors enhances further cancer-selective activity of the GAPE-Prom. The GAPE-Prom provides a genetic tool to specifically regulate transgene expression in cancer cells.
Document Details
- Document Type
- Pub Defense Publication
- Publication Date
- Mar 15, 2022
- Source ID
- 10.3390/cancers14061497
Entities
People
- Amit Kumar
- Anjan K. Pradhan
- Devanand Sarkar
- Dipankar Bandyopadhyay
- Joseph W Landry
- Luni Emdad
- Padmanabhan Mannangatti
- Paul B Fisher
- Praveen Bhoopathi
- Santanu Maji
- Swadesh K. Das
- Xiang-Yang Wang
- Xiaoyan Deng
Organizations
- National Foundation for Cancer Research
- National Institutes of Health
- United States Department of Defense