TATk-CDKL5 fusion proteins, compositions, formulations, and use thereof
Disclosed herein are compositions and formulations containing a TATk-CDKL5 fusion protein. Also disclosed are methods of producing a TATk-CDKL5 fusion protein from vectors containing a TATk-CDKL5 cDNA and methods of transducing cells with the vectors containing a TATk-CDKL5 cDNA and the TATk-CDKL5 fusion protein.
1. A recombinant nucleic acid comprising:
a CDKL5 nucleic acid sequence, wherein the CDKL5 nucleic acid sequence encodes a CDKL5 polypeptide sequence, wherein the CDKL5 polypeptide sequence comprises SEQ ID NO: 2 or SEQ ID NO: 16; and
a TATk nucleic acid sequence, wherein the TATk nucleic acid sequence encodes a TATk polypeptide sequence, wherein the TATk polypeptide sequence has 90% to 100% sequence identity to SEQ ID NO: 4 and wherein the TATk nucleic acid sequence is operatively coupled to the CDKL5 nucleic acid sequence and/or the TATk polypeptide sequence is operatively coupled to the CDKL5 polypeptide sequence.
2. The recombinant nucleic acid of claim 1 , wherein the CDKL5 nucleic acid has 90% to 100% sequence identity to SEQ ID NO: 1 or SEQ ID NO: 15.
3. The recombinant nucleic acid of claim 1 , wherein the TATk nucleic acid sequence has 90% to 100% sequence identity to SEQ ID NO: 3.
4. The recombinant nucleic acid of claim 1 , wherein the nucleic acid encodes a fusion protein that increases neurite growth, elongation, branch number, or branch density in the brain of a subject as compared to a control.
5. The recombinant nucleic acid of claim 1 , wherein the nucleic acid encodes a fusion protein that reduces neuron apoptosis in the brain of a subject as compared to a control.
6. The recombinant nucleic acid of claim 1 , wherein the recombinant nucleic acid further comprises a nucleic acid sequence encoding a protein tag, wherein the nucleic acid sequence encoding the protein tag is operatively coupled to the CDKL5 nucleic acid sequence.
7. The recombinant nucleic acid of claim 6 , wherein the protein tag is selected from the group consisting of: chitin binding protein (CBP), maltose binding protein (MBP), glutathione-S-transferase (GST), poly(His), thioredoxin (TRX), poly(NANP), a FLAG-tag, V5-tag, Myc-tag, HA-tag, S-tag, SBP-Tag, Sftag 1, Softag 3, Tc tag, Xpress tag, Strep-tag, Isopeptag, Spy Tag, Ty tag, Biotin Carboxyl Carrier Protein (BCCP), and Nus tag.
8. The recombinant nucleic acid of claim 1 , wherein the recombinant nucleic acid further comprises a nucleic acid sequence encoding a reporter protein, wherein the nucleic acid sequence encoding the reporter protein is operatively coupled to the CDKL5 nucleic acid sequence.
9. The recombinant nucleic acid of claim 8 , wherein the reporter protein is selected from the group consisting of: a fluorescent protein, beta-galactosidase, a luciferase protein, an antibiotic-resistance protein, p-glucuronidase, and alkaline phosphatase.
10. A vector comprising the recombinant nucleic acid of claim 1 .
11. The vector of claim 10 , wherein the vector is suitable for expression in a bacterial cell.
12. The vector of claim 10 , wherein the vector is suitable for expression in a mammalian cell.
13. A method comprising transfecting cells with the recombinant nucleic acid of claim 1 .
14. The method of claim 13 , wherein the recombinant nucleic acid is contained in a vector.
15. The method of claim 13 , wherein the cells comprise bacterial cells.
16. The method of claim 13 , wherein the cells comprise mammalian cells.