Isolation and identification of mouse and human transcription control elements associated with cytochrome expression
View Patent ↗The present invention relates to transcription control elements derived from mouse and human genes associated with cytochrome expression, e.g., Cyp3A11 and CYP3A4, respectively. Isolated polynucleotides, expression cassettes, vectors, recombinant cells, and transgenic animals, may comprise such transcription control elements as described herein.
1. An expression cassette that activates expression of a reporter sequence in mouse liver cells, the expression cassette comprising
a reporter sequence; and
a polynucleotide comprising an inducible transcriptional control element and having at least 95% identity to nucleotides 1-11,002 of SEQ ID NO:12,
wherein the polynucleotide is operably linked to the reporter sequence such that when expression of the polynucleotide is induced by a compound that activates expression of the reporter sequence, the reporter sequence is expressed in liver cells.
2. The expression cassette of claim 1 , wherein the reporter sequence encodes a light-generating protein.
3. The expression cassette of claim 2 , wherein the light-generating protein is a bioluminescent protein or a fluorescent protein.
4. The expression cassette of claim 3 , wherein the bioluminescent protein is luciferase.
5. The expression cassette of claim 3 , wherein the fluorescent protein is selected from the group consisting of blue fluorescent protein, cyan fluorescent protein, green fluorescent protein, yellow fluorescent protein, and red fluorescent protein.
6. A vector comprising
(a) the expression cassette of claim 1 ; and
(b) a vector backbone.
7. The vector of claim 6 , wherein said vector backbone further comprises a selectable marker.
8. A mouse liver cell comprising the expression cassette of claim 1 .
9. A mouse liver cell comprising a vector of claim 6 .
10. A transgenic mouse comprising liver cells, wherein the liver cells comprise the expression cassette of claim 1 .
11. A method for identifying an analyte that modulates expression of a sequence encoding a light-generating protein, in a living mouse, said method comprising
intravenously administering to a living mouse via the tail vein a vector mixture comprising the expression cassette of claim 2 such that hepatocytes of the mouse express the light-generating protein in the presence of a compound that activates expression of the sequence encoding the light-generating protein,
administering to said mouse said analyte; and
monitoring expression of said light-generating protein, wherein expression of the light-generating protein indicates that the analyte modulates expression of the reporter sequence.
12. A method for monitoring expression of a reporter sequence in an isolated mouse liver cell, said method comprising:
monitoring, in the presence of a compound that activates expression of the reporter sequence, the expression of a reporter sequence in an isolated mouse liver cell, said cell comprising the expression cassette of claim 1 .