AAV virions with decreased immunoreactivity and uses therefor
Methods of making and using recombinant AAV virions with decreased immunoreactivity are described. The recombinant AAV virions include mutated capsid proteins or are derived from non-primate mammalian AAV serotypes and isolates that display decreased immunoreactivity relative to AAV-2.
1. A mutated adeno-associated virus serotype 2 (AAV-2) capsid protein comprising a substitution of one or more of the amino acids occurring at a position corresponding to a position of the full-length AAV-2 VP2 capsid,
wherein the one or more amino acids comprise amino acid 449 substituted with a lysine or an alanine, and
wherein the mutated AAV-2 capsid protein displays an increase in resistance to neutralization by one or more human IgGs as compared to the wild-type AAV-2 capsid protein.
2. The mutated AAV-2 capsid protein of claim 1 , wherein the one or more amino acids are selected from the group consisting of amino acid:
334 substituted with an alanine and 449 substituted with an alanine;
334 substituted with an alanine, 360 substituted with a lysine, 394 substituted with an alanine, 411 substituted with an alanine, 413 substituted with an alanine, and 449 substituted with an alanine;
334 substituted with an alanine, 360 substituted with a lysine, 449 substituted with a lysine, and 568 substituted with a lysine;
334 substituted with an alanine, 360 substituted with a lysine, and 449 substituted with a lysine;
334 substituted with an alanine, 449 substituted with a lysine, and 568 substituted with a lysine;
334 substituted with an alanine, 449 substituted with an alanine, and 568 substituted with an alanine;
347 substituted with a cysteine, 449 substituted with an alanine, and 450 substituted with an alanine;
411 substituted with an alanine, 413 substituted with an alanine, 449 substituted with an alanine, and 450 substituted with an alanine;
411 substituted with an alanine, 413 substituted with an alanine, 449 substituted with an alanine, 450 substituted with an alanine, 568 substituted with an alanine, and 571 substituted with an alanine;
449 substituted with a lysine and 568 substituted with a lysine; and
449 substituted with an alanine and 450 substituted with an alanine.
3. A polynucleotide encoding the mutated AAV-2 capsid protein of claim 1 .
4. A polynucleotide encoding the mutated AAV-2 capsid protein of claim 2 .
5. A recombinant AAV virion comprising the mutated AAV-2 capsid protein of claim 1 .
6. A recombinant AAV virion comprising the mutated AAV-2 capsid protein of claim 2 .
7. A method of delivering a recombinant AAV virion to a cell or tissue of a vertebrate subject, said method comprising:
(a) providing a recombinant AAV virion according to claim 5 ;
(b) delivering said recombinant AAV virion to said cell or tissue.
8. A method of delivering a recombinant AAV virion to a cell or tissue of a vertebrate subject, said method comprising:
(a) providing a recombinant AAV virion according to claim 6 ;
(b) delivering said recombinant AAV virion to said cell or tissue.
9. The mutated AAV-2 capsid protein of claim 1 , wherein the one or more amino acids are amino acid 334 substituted with an alanine and amino acid 449 substituted with an alanine.
10. The mutated AAV-2 capsid protein of claim 1 , wherein the one or more amino acids are amino acid 334 substituted with an alanine, amino acid 449 substituted with a lysine, and amino acid 568 substituted with a lysine.
11. The mutated AAV2 capsid protein of claim 1 , wherein the mutated AAV2 capsid protein is a VP2 capsid protein.
12. The mutated AAV2 capsid protein of claim 1 , wherein the one or more amino acids comprise amino acid 449 substituted with a lysine.
13. The mutated AAV2 capsid protein of claim 1 , wherein the one or more amino acids comprise amino acid 449 substituted with an alanine.
14. The recombinant AAV virion of claim 5 , wherein said virion comprises a heterologous nucleic acid molecule encoding an antisense RNA or a ribozymes.
15. The recombinant AAV virion of claim 5 , wherein said virion comprises a heterologous nucleic acid molecule encoding a therapeutic protein operably linked to control elements capable of directing the in vivo transcription and translation of said protein.