Adeno-associated virus (AAV) systems for treatment of genetic hearing loss
Described herein are codon optimized nucleic acid sequences encoding gap junction protein beta 2 (GJB2), and associated genetic elements, for use in recombinant adeno-associated virus (rAAV)-based gene therapy. The rAAV vectors through which GJB2 is packaged can be used for targeted delivery to patients suffering from genetic hearing loss, including patients with autosomal mutations, recessive or dominant, in the GJB2 gene.
1. An isolated polynucleotide comprising a nucleic acid sequence encoding a codon optimized gap junction protein beta 2 (GJB2), wherein the nucleic acid sequence is at least 85% identical to SEQ ID NO: 18.
2. The polynucleotide of claim 1 , comprising in the following order CBA-GJB2 (X)-HA-WPRE-pA, where X comprises a nucleic acid sequence at least 85% identical to SEQ ID NO: 18.
3. A host cell comprising the polynucleotide of claim 1 .
4. A recombinant herpes simplex virus (rHSV) comprising the polynucleotide of claim 1 .
5. A transgene expression cassette comprising:
the polynucleotide of claim 1 ; and
minimal regulatory elements.
6. A nucleic acid vector comprising the expression cassette of claim 5 .
7. The vector of claim 6 , wherein the vector is an adeno-associated viral (AAV) vector.
8. A host cell comprising the transgene expression cassette of claim 5 .
9. A composition comprising the host cell of claim 8 .
10. A composition comprising the transgene expression cassette of claim 5 .
11. A recombinant adeno-associated (rAAV) expression vector comprising the polynucleotide of claim 1 and an AAV genomic cassette.
12. The expression vector of claim 11 , wherein the AAV genomic cassette is flanked by two sequence-modulated inverted terminal repeats.
13. The expression vector of claim 11 further comprising a protein capsid variant optimally suited for cochlear delivery.
14. A composition comprising the rAAV expression vector of claim 11 .
15. A composition comprising the polynucleotide of claim 1 .
16. The polynucleotide of claim 1 , wherein the nucleic acid sequence encoding the codon optimized GJB2 protein is at least 95% identical to SEQ ID NO: 18.
17. The polynucleotide of claim 1 , wherein the nucleic acid sequence encoding the codon optimized GJB2 protein is at least 99% identical to SEQ ID NO: 18.
18. The polynucleotide of claim 1 , wherein the nucleic acid sequence encoding the codon optimized GJB2 protein consists of SEQ ID NO: 18.