Development of ligand-dependent regulatory system and its application to gene therapy of insulin-dependent diabetes mellitus.
- Author:
Jong Min KIM
1
;
Soo Jin KIM
;
Hyun Chul LEE
;
Kyung Sup KIM
Author Information
1. Research Center for Ischemic Tissue Regeneration and Medical Research Institute, College of Medicine, Pusan National University, Busan 602-739, Korea. jhkimst@pusan.ac.kr
- Publication Type:Original Article ; Evaluation Studies ; Research Support, Non-U.S. Gov't
- Keywords:
apoptosis;
lysophosphatidylcholines;
neu-rites;
PC12 cells;
phospholipase D2
- MeSH:
Transfection;
Trans-Activation (Genetics);
Receptors, Cytoplasmic and Nuclear/genetics;
Mice, Inbred BALB C;
Mice;
Male;
*Ligands;
Genetic Vectors/*chemical synthesis;
Genes, Reporter;
Gene Therapy/*methods;
*Gene Expression Regulation;
Diabetes Mellitus, Type 1/*therapy;
Diabetes Mellitus, Experimental/blood/*therapy;
Blood Glucose/analysis;
Animals
- From:Experimental & Molecular Medicine
2006;38(4):385-392
- CountryRepublic of Korea
- Language:English
-
Abstract:
To develop an inducible expression system, the enhanced artificial nuclear receptors and target reporters were constructed. Artificial nuclear receptors were generated by fusing three domains, consisting of DNA-binding domain (DBD) of GAL4, ligand binding domain (LBD) of progesterone or estrogen receptor, and activation domain (AD) of VP16, sterol regulatory element binding protein (SREBP)-1a, or SREBP-2. The activation domain of SREBP-1a showed most potent transcriptional activity. The maximal level of target reporter gene expression was extremely elevated by the usage of ATP citrate-lyase (ACL) minimal promoter -60/+67 in place of artificial TATA promoter, while the SV40 enhancer severely increased the basal transcription in the absence of ligand. The induction system, developed in the present study, was applied to cell therapy, resulting in successful induction of single-chain insulin analogue (SIA) gene expression to correct the hyperglycemia in diabetic animals. By means of subcutaneous cell therapy, the SIA gene expression rapidly occurred after the local topical application of ligand. These results suggest that our system represents a powerful tool for transcriptional regulation of target gene that can be used for diverse applications, ranging from basic research to gene therapy.
