Insulators prevent transcriptional interference between two promoters in a double gene construct for transgenesis

• Published in: FEBS letters Vol. 520; no. 1; pp. 47 - 52
• Main Authors: Hasegawa, Kouichi, Nakatsuji, Norio
• Format: Journal Article
• Language: English
• Published: England Elsevier B.V 05.06.2002
• Subjects: Animals; Bacterial Proteins - genetics; Bacterial Proteins - metabolism; Binding Sites - genetics; CAG, cytomegalovirus enhancer and chicken β-actin promoter; Cell Line; Chickens; Double gene transfection; Enhancer blocking; Female; Fluorescent protein; Gene Expression Regulation; Gene Silencing; GFP, green fluorescent protein; Globins - genetics; Green Fluorescent Proteins; Humans; Insulator; Insulin - genetics; Insulin - metabolism; Luminescent Proteins - genetics; Luminescent Proteins - metabolism; Male; Mice; Mice, Transgenic; Microscopy, Fluorescence; Promoter Regions, Genetic - genetics; Rats; Recombinant Fusion Proteins - genetics; Recombinant Fusion Proteins - metabolism; Red Fluorescent Protein; RFP, red fluorescent protein; Transcription, Genetic - genetics; Transcriptional interference; Transfection; Transgenes - genetics; Transgenic mouse; YFP, yellow fluorescent protein; GFP, green fluorescent protein; Enhancer blocking; Insulator; Double gene transfection; YFP, yellow fluorescent protein; Fluorescent protein; Transcriptional interference; CAG, cytomegalovirus enhancer and chicken β-actin promoter; Transgenic mouse; RFP, red fluorescent protein
• ISSN: 0014-5793; 1873-3468
• DOI: 10.1016/S0014-5793(02)02761-8

In transgenesis, the expression of two transgenes is often subject to mutual interference by each of the two expression cassettes when they are driven by different transcriptional regulatory elements in a single construct. To study this problem, we constructed vectors consisting of two expression units, one contains a strong ubiquitous promoter and the other contains a tissue-specific transcriptional element. The expression pattern of each transgene was examined in transfected cell lines and also in transgenic mice. In both cases, two expression units in a single construct were expressed in an independent manner and were controlled by their respective regulatory element only if we placed insulators at both ends of one expression unit. These results indicate that usage of insulators is a valuable tool for transfection of double gene constructs in transgenesis.