Methods and compositions for regulation of transgene expression
Nucleases and methods of using these nucleases for expressing a transgene from a safe harbor locus in a secretory tissue, and clones and animals derived therefrom.
1. A polynucleotide encoding a non-naturally occurring fusion protein comprising
(i) a Transcription Activator Like Effector (TALE) DNA binding domain that binds to an endogenous albumin gene, wherein the TALE DNA binding protein comprises a plurality of TALE repeat units, each TALE repeat unit comprising an amino acid Repeat Variable Diresidue (RVD) that binds to a nucleotide in a target sequence in an endogenous albumin gene, wherein the TALE DNA binding domain comprises a +17 or +63C-terminal truncation and
(ii) a cleavage domain,
wherein the non-naturally occurring fusion protein cleaves the endogenous albumin gene.
2. An isolated cell comprising one or more polynucleotides according to claim 1 .
3. The isolated cell of claim 2 , wherein the cell is a stem cell.
4. The cell of claim 3 , wherein the stem cell is selected from the group consisting of an embryonic stem cell (ESC), an induced pluripotent stem cell (iPSC), a hepatic stem cell and a liver stem cell.
5. A kit comprising the polynucleotide according to claim 1 .
6. A method of cleaving an endogenous albumin gene in a cell, the method comprising:
introducing, into the cell, one or more polynucleotides according to claim 1 , under conditions such that the one or more fusion proteins—are expressed and the endogenous albumin gene is cleaved.
7. The method of claim 6 , wherein the one or more polynucleotides comprise mRNA.
8. The method of claim 6 , wherein the one or more polynucleotides are included in one or more expression vectors.
9. The method of claim 6 , wherein the cell is a liver cell.