Methods for using transcription-dependent directed evolution of AAV capsids
Disclosed are methods for performing transcription-dependent directed evolution (TRADE) and novel AAV capsids selected using such methods.
1. An adeno-associated virus serotype 9 (AAV9) capsid protein comprising an amino acid insertion within loop 8, wherein the amino acid insertion comprises SEQ ID NO: 128, and wherein the AAV9 capsid protein without the amino acid insertion comprises 90% sequence identity to SEQ ID NO: 221.
2. The AAV9 capsid protein of claim 1 , wherein the AAV9 capsid protein comprises a N272A mutation.
3. The AAV9 capsid protein of claim 1 , wherein the AAV9 capsid protein comprises SEQ ID NO: 221.
4. The AAV9 capsid protein of claim 1 , wherein the AAV9 capsid protein comprises SEQ ID NO: 222.
5. The AAV9 capsid protein of claim 1 wherein the amino acid insertion within loop 8 comprises a substitution of position 588 of SEQ ID NO: 221 with the amino acid insertion.
6. The AAV9 capsid protein of claim 1 , wherein the amino acid insertion further comprises a linker.
7. A nucleic acid molecule encoding an adeno-associated virus serotype 9 (AAV9) capsid protein comprising an amino acid insertion within loop 8 of the AAV9 capsid protein, wherein the amino acid insertion comprises SEQ ID NO: 128, and wherein the AAV9 capsid protein without the amino acid insertion comprises 90% sequence identity to SEQ ID NO: 221.
8. The nucleic acid molecule of claim 7 , wherein the AAV9 capsid protein comprises a N272A mutation.
9. The nucleic acid molecule of claim 7 , wherein the AAV9 capsid protein comprises SEQ ID NO: 221.
10. The nucleic acid molecule of claim 7 , wherein the AAV9 capsid protein comprises SEQ ID NO: 222.
11. The nucleic acid molecule of claim 7 , wherein the amino acid insertion within loop 8 comprises a substitution of position 588 of SEQ ID NO: 221 with the amino acid insertion.
12. The nucleic acid molecule of claim 7 , wherein the amino acid insertion further comprises a linker.