US 11,993,777 B2
Compositions and methods for treating non-age-associated hearing impairment in a human subject
Emmanuel John Simons, Brookline, MA (US); Ellen Reisinger, Dußlingen (DE); Sebastian Kügler, Göttingen (DE); and Hanan Al-Moyed, Göttingen (DE)
Assigned to Akouos, Inc., Boston, MA (US)
Filed by Akouos, Inc., Boston, MA (US)
Filed on Aug. 30, 2023, as Appl. No. 18/458,744.
Application 18/458,744 is a continuation of application No. 17/929,647, filed on Sep. 2, 2022, granted, now 11,781,145.
Application 17/929,647 is a continuation of application No. 17/378,606, filed on Jul. 16, 2021, granted, now 11,525,139, issued on Dec. 13, 2022.
Application 17/378,606 is a continuation of application No. 16/327,396, abandoned, previously published as PCT/US2017/048257, filed on Aug. 23, 2017.
Claims priority of provisional application 62/494,866, filed on Aug. 23, 2016.
Prior Publication US 2023/0407315 A1, Dec. 21, 2023
This patent is subject to a terminal disclaimer.
Int. Cl. C12N 15/52 (2006.01); A01K 67/027 (2006.01); A01K 67/0276 (2024.01); A61K 38/17 (2006.01); A61K 48/00 (2006.01); A61P 27/16 (2006.01); C07K 14/47 (2006.01); C12N 5/0793 (2010.01); C12N 9/16 (2006.01); C12N 15/65 (2006.01); C12N 15/86 (2006.01); C12N 15/90 (2006.01)
CPC C12N 15/52 (2013.01) [A01K 67/0276 (2013.01); A61K 38/1709 (2013.01); A61K 48/0075 (2013.01); A61P 27/16 (2018.01); C07K 14/47 (2013.01); C12N 5/062 (2013.01); C12N 9/16 (2013.01); C12N 15/65 (2013.01); C12N 15/86 (2013.01); C12N 15/902 (2013.01); A01K 2217/075 (2013.01); A01K 2227/103 (2013.01); A01K 2267/03 (2013.01); C12N 2310/20 (2017.05); C12N 2750/14143 (2013.01); C12N 2830/008 (2013.01); C12Y 301/03001 (2013.01)] 29 Claims
 
1. A composition comprising two different nucleic acid vectors, wherein:
each of the two different vectors comprises a coding sequence that encodes a different portion of an otoferlin protein, each of the encoded portions being at least 30 amino acid residues in length;
no single vector of the at least two different vectors encodes a full-length otoferlin protein;
a first of the two different vectors comprises a promoter operably linked to a coding sequence that encodes an N-terminal portion of the otoferlin protein;
at least one of the coding sequences comprises a nucleotide sequence spanning two neighboring exons of otoferlin genomic DNA, and lacks an intronic sequence between the two neighboring exons; and
when introduced into a mammalian cell the at least two different vectors undergo concatamerization or homologous recombination with each other, thereby forming a recombined nucleic acid that encodes a full-length otoferlin protein.
 
16. A composition comprising two different nucleic acid vectors, wherein:
a first nucleic acid vector of the two different nucleic acid vectors comprises a promoter, a first coding sequence that encodes an N-terminal portion of an otoferlin protein positioned 3′ of the promoter, and a splicing donor signal sequence positioned at the 3′ end of the first coding sequence; and
a second nucleic acid vector of the two different nucleic acid vectors comprises a splicing acceptor signal sequence, a second coding sequence that encodes a C-terminal portion of an otoferlin protein positioned at the 3′ end of the splicing acceptor signal sequence, and a polyadenylation sequence at the 3′ end of the second coding sequence;
wherein each of the encoded portions is at least 30 amino acid residues in length,
wherein the amino acid sequences of the encoded portions do not overlap,
wherein no single vector of the two different vectors encodes a full-length otoferlin protein, and,
when the coding sequences are transcribed in a mammalian cell, to produce RNA transcripts, splicing occurs between the splicing donor signal sequence on one transcript and the splicing acceptor signal sequence on the other transcript, thereby forming a recombined RNA molecule that encodes a full-length otoferlin protein.