Generating mammalian T cell activation inducible synthetic promoters (SYN+PRO) to improve T cell therapy
Aspects of the invention described herein relate to methods of making and using inducible promoters for transgene expression. The inducible promoters are derived from the NFAT-RE inducible system and are used to improve or enhance T cell survival and proliferation.
1. An isolated nucleic acid comprising a promoter comprising a transcription response element having a sequence with at least 98% sequence identity to the sequence as set forth in SEQ ID NO: 7.
2. The isolated nucleic acid of claim 1 , wherein the transcription response element comprises a sequence having at least 99% sequence identity to the sequence as set forth in SEQ ID NO: 7.
3. The isolated nucleic acid of claim 1 , wherein the transcription response element comprises the sequence as set forth in SEQ ID NO: 7.
4. The isolated nucleic acid of claim 1 , wherein the promoter further comprises an IL2 minimal promoter.
5. The isolated nucleic acid of claim 1 , wherein the isolated nucleic acid is operably linked to a sequence encoding a payload.
6. The isolated nucleic acid of claim 5 , wherein the payload comprises CD122, CD127, CD360, caSTAT5, dnSHP1, or dnSHP2; PD1:MyD88, PD1:CD28, CD200:CD28, or miRNA155.
7. A vector comprising a nucleic acid comprising a promoter comprising a transcription response element having a sequence with at least 98% sequence identity to the sequence as set forth in SEQ ID NO: 7,
wherein the nucleic acid is operably linked to a sequence encoding a payload.
8. The vector of claim 7 , wherein the transcription response element comprises a sequence having at least 99% sequence identity to the sequence as set forth in SEQ ID NO: 7.
9. The vector of claim 7 , wherein the transcription response element comprises the sequence as set forth in SEQ ID NO: 7.
10. The vector of claim 7 , wherein the promoter further comprises a minimal IL2 promoter.
11. The vector of claim 7 , wherein the vector comprises a viral vector, a transposase based minicircle, or a nanoplasmid.
12. The vector of claim 7 , wherein the vector comprises a lentiviral vector.
13. The vector of claim 7 , wherein the payload comprises CD122, CD127, CD360, caSTAT5, dnSHP1, or dnSHP2; PD1:MyD88, PD1:CD28, CD200:CD28, or miRNA155.
14. A cell comprising: the vector of claim 7 .
15. The cell of claim 14 , wherein the transcription response element comprises a sequence having at least 99% sequence identity to the sequence as set forth in SEQ ID NO: 7.
16. The cell of claim 14 , wherein the transcription response element comprises the sequence as set forth in SEQ ID NO: 7.
17. The cell of claim 14 , wherein the promoter further comprises a minimal IL2 promoter.
18. The cell of claim 14 , wherein the cell is a T cell or a hematopoietic stem cell.
19. The cell of claim 18 , wherein the T cell is a CD4+ T cell or a CD8+ T cell.
20. The cell of claim 14 , wherein the cell further comprises a nucleic acid encoding a chimeric antigen receptor (CAR).