LexA-based chemical-inducible gene expression system for use in transgenic animals

摘要

The present invention relates generally to chemical-inducible system and to methods of use in transgenic animals. More specifically, the present invention relates to a chimeric transcription factor that binds to a ligand and functions in ligand-dependent manner to induce expression of genes of interest under the control of a synthetic operator-promoter sequence. The expression of genes of interest can be tightly controlled by adding or removing the ligand.

主权项

1. A system useful for preparing a non-human transgenic animal which comprises a first nucleic acid molecule comprising a first DNA construct and a second DNA construct, wherein:
(a) the first DNA construct encoding a chimeric transcription factor and having the following elements: (i) a DNA encoding a DNA binding domain of a bacterial LexA protein, (ii) a DNA encoding a truncated ligand binding domain of a human progesterone receptor and (iii) a DNA encoding an activation domain of the p65 subunit of human NF-κB, wherein the DNA of (i) comprises the nucleotide sequence set forth in SEQ ID NO:6 and the DNA of (ii) and (iii) comprises the nucleotide sequence set forth in SEQ ID NO:9; and wherein (b) the second DNA construct has the following elements in the 5′ to 3′ direction: (i) a operator comprising LexA binding sites, (ii) a promoter and (iii) a gene, wherein the operator comprises the nucleotide sequence set forth in SEQ ID NO:15.