TCR(alpha)-LCR-derived gene regulatory cassettes
View Patent ↗A method of delivering a transgene to a cell is provided. The method uses a vector that contains a T-cell receptor alpha locus control region (TCRαLCR) derived gene regulatory cassette having fewer than 5.0-kb. The method delivers the transgene with spatiotemporally specific gene expression and silencing-prevention controls to a cell such that a predetermined subset of progeny cell-types express a gene product from the transgene. Other progeny of the cell diminish, or silence, the expression of the gene product.
1. A method of delivering a gene to a cell such that progeny of the cell expresses a predetermined gene product with spatiotemporal specificity and resistance to silencing, the method comprising:
introducing, in an in vitro environment, a vector to a cell, the vector comprising:
(a) a T-cell receptor alpha locus control region (TCRaLCR) derived gene regulatory cassette having fewer than 5.0-kb nucleotides, the TCRaLCR derived gene regulatory cassette comprising a sequence that is SEQ. ID NO: 1 and
(b) a first transgene to be expressed.
2. The method as recited in claim 1 , wherein the cell is a stem cell.
3. The method as recited in claim 1 , wherein the cell is a multi-potent precursor cell.
4. The method as recited in claim 3 , further comprising permitting the multi-potent precursor cell to proliferate and differentiate to form a population of T-cells and a population of non-T-cell progeny cells, wherein the population of T-cells differentiating from the multi-potent precursor cell expresses the first transgene with a first degree of expression and the population of non-T-cell progeny cells expresses the first transgene with a second degree of expression that is lower than the first degree of expression.
5. The method as recited in claim 4 , wherein the second degree of expression is zero.
6. The method as recited in claim 1 , wherein the first transgene is directly linked 5′ to the TCRαLCR derived gene regulatory cassette.
7. The method as recited in claim 1 , wherein the first transgene is directly linked 3′ to the TCRαLCR derived gene regulatory cassette.
8. The method as recited in claim 1 , wherein the first transgene is promoter-driven cDNA.
9. The method as recited in claim 1 , wherein the vector further comprises a second transgene, wherein first transgene and the second transgene flank the T-cell receptor alpha locus control region (TCRαLCR) derived gene regulatory cassette.
10. The method as recited in claim 1 , wherein the vector further comprises a second transgene and the second transgene is a promoter-driven cDNA.
11. The method as recited in claim 1 , wherein the vector consists of the T-cell receptor alpha locus control region (TCRaLCR) derived gene regulatory cassette and the first transgene.
12. The method as recited in claim 1 , wherein the vector consists of the T-cell receptor alpha locus control region (TCRaLCR) derived gene regulatory cassette and the first transgene and a second transgene.
13. A vector for delivering a gene to a cell such that progeny of the cell expresses a predetermined gene product with spatiotemporal specificity and resistance to silencing, the vector comprising:
(a) a T-cell receptor alpha locus control region (TCRaLCR) derived gene regulatory cassette having fewer than 5.0-kb nucleotides, the TCRaLCR derived gene regulatory cassette comprising, a sequence that is SEQ ID NO: 1 and
(b) a first transgene to be expressed.
14. The vector as recited in claim 13 , wherein the vector further comprises a second transgene, wherein first transgene and the second transgene flank the T-cell receptor alpha locus control region (TCRαLCR) derived gene regulatory cassette.
15. The vector as recited in claim 13 , wherein the vector further comprises a second transgene and the second transgene is a promoter-driven cDNA.