Dept. of Cell and Developmental Biology
Oregon Health & Science University
Understanding mechanisms of gene regulation has broad therapeutic implications for human disease. Here we describe a novel method for generating human cell lines that serve as reporters of transcriptional activity. This method exploits the ability of recombinant adena-associated virus (rAA V) to mediate the insertion of exogenous DNA sequences into specific genomic loci through homologous recombination. To overcome the severe size limitation of the rAAV for carrying exogenous DNA, an EGFP-Luciferase fusion gene was used as both a selectable marker and gene expression reporter. EGFP was used for selection of correctly targeted alleles by taking advantage of known regulatory conditions that activate transcription of specific genes. Using this method, we describe the generation of primary human fibroblasts that express EGFP-Luciferase under the control of the c-Myc oncogene.
School of Medicine
Fernandez, Sandra L., "Development of novel human reporter cell lines using recombinant adeno-associated virus mediated homologous recombination" (2007). Scholar Archive. 802.