US 12,275,959 B2
Compositions and methods for enhanced gene expression in cone cells
Thomas W. Chalberg, Jr., Redwood City, CA (US); Jay Neitz, Seattle, WA (US); and Maureen Neitz, Seattle, WA (US)
Assigned to Adverum Biotechnologies, Inc., Redwood City, CA (US)
Filed by Adverum Biotechnologies, Inc., Redwood City, CA (US); and University of Washington, Seattle, WA (US)
Filed on Dec. 20, 2021, as Appl. No. 17/556,847.
Application 17/556,847 is a continuation of application No. 15/984,085, filed on May 18, 2018, granted, now 11,248,214.
Application 15/984,085 is a continuation of application No. 14/660,657, filed on Mar. 17, 2015, granted, now 10,000,741, issued on Jun. 19, 2018.
Claims priority of provisional application 62/127,185, filed on Mar. 2, 2015.
Claims priority of provisional application 61/954,330, filed on Mar. 17, 2014.
Prior Publication US 2022/0259568 A1, Aug. 18, 2022
Int. Cl. C12N 7/00 (2006.01); A61K 35/761 (2015.01); A61K 48/00 (2006.01); C12N 15/86 (2006.01); A61K 38/00 (2006.01)
CPC C12N 7/00 (2013.01) [A61K 35/761 (2013.01); C12N 15/86 (2013.01); A61K 38/00 (2013.01); A61K 48/00 (2013.01); C12N 2750/14143 (2013.01); C12N 2800/24 (2013.01); C12N 2830/008 (2013.01); C12N 2830/30 (2013.01); C12N 2830/42 (2013.01)] 16 Claims
 
1. A polynucleotide cassette for enhanced expression of a transgene in cone cells of a mammalian retina, comprising:
(a) a promoter region consisting of a truncated opsin promoter having a sequence selected from the group consisting of (i) 95% or more sequence identity to SEQ ID NO: 55 or functional fragment thereof comprising SEQ ID NO:80, (ii) SEQ ID NO: 82, and (iii) SEQ ID NO: 83;
(b) a coding sequence operatively linked to the promoter region; and
(c) a polyadenylation site operatively linked to the coding sequence.
 
14. A pharmaceutical composition comprising an rAAV comprising (a) an AAV capsid protein and (b) a polynucleotide cassette for enhanced expression of a transgene in cone cells of a mammalian retina, comprising:
(a) a promoter region consisting of a truncated opsin promoter having a sequence selected from the group consisting of (i) 95% or more sequence identity to SEQ ID NO: 55 or functional fragment thereof comprising SEQ ID NO:80, (ii) SEQ ID NO: 82, and (iii) SEQ ID NO: 83;
(b) a coding sequence operatively linked to the promoter region; and
(c) a polyadenylation site operatively linked to the coding sequence; wherein the polynucleotide cassette is flanked by AAV ITRs and a pharmaceutical excipient.
 
15. A method for expressing a transgene in cone cells, comprising:
contacting one or more cone cells with an effective amount of a recombinant adeno-associated virus comprising (a) an AAV capsid protein and (b) a polynucleotide cassette for enhanced expression of a transgene in cone cells of a mammalian retina, comprising:
(a) a promoter region consisting of a truncated opsin promoter having a sequence selected from the group consisting of (i) 95% or more sequence identity to SEQ ID NO: 55 or functional fragment thereof comprising SEQ ID NO:80, (ii) SEQ ID NO: 82, and (iii) SEQ ID NO: 83;
(b) a coding sequence operatively linked to the promoter region; and
(c) a polyadenylation site operatively linked to the coding sequence; wherein the polynucleotide cassette is flanked by AAV ITRs, wherein the transgene is expressed at detectable levels in the one or more cone cells.