US 12,441,994 B2
Methods for targeted insertion of DNA in genes
Nicholas J. Baltes, Oakdale, MN (US)
Assigned to BLUEALLELE CORPORATION, Oakdale, MN (US)
Filed by BLUEALLELE CORPORATION, Oakdale, MN (US)
Filed on Dec. 1, 2023, as Appl. No. 18/526,758.
Application 18/526,758 is a continuation of application No. 17/830,011, filed on Jun. 1, 2022, granted, now 12,054,706.
Application 17/830,011 is a continuation of application No. 17/590,613, filed on Feb. 1, 2022, granted, now 11,365,407, issued on Jun. 1, 2022.
Application 17/590,613 is a continuation of application No. 17/366,290, filed on Jul. 2, 2021, granted, now 11,254,930, issued on Feb. 2, 2022.
Application 17/366,290 is a continuation of application No. 16/800,444, filed on Feb. 25, 2020, granted, now 11,091,756, issued on Jul. 28, 2021.
Application 16/800,444 is a continuation of application No. 16/601,144, filed on Oct. 14, 2019, abandoned.
Claims priority of provisional application 62/864,432, filed on Jun. 20, 2019.
Claims priority of provisional application 62/830,654, filed on Apr. 8, 2019.
Claims priority of provisional application 62/746,497, filed on Oct. 16, 2018.
Prior Publication US 2024/0141319 A1, May 2, 2024
This patent is subject to a terminal disclaimer.
Int. Cl. C12N 15/10 (2006.01); C12N 15/86 (2006.01); C12N 15/90 (2006.01)
CPC C12N 15/102 (2013.01) [C12N 15/86 (2013.01); C12N 15/907 (2013.01); C12N 2310/20 (2017.05); C12N 2800/80 (2013.01)] 19 Claims
 
1. A vector comprising a transgene comprising in 5′ to 3′ orientation: a first splice acceptor, a first coding sequence, a first terminator, a second terminator reverse complement, a second coding sequence reverse complement, and a second splice acceptor reverse complement,
wherein the first coding sequence is operably linked to the first splice acceptor and first terminator, and the second coding sequence is operably linked to the second splice acceptor and second terminator, wherein the first terminator is selected from an SV40 poly (A) or BGH poly (A),
wherein the first and second coding sequences differ in nucleic acid sequence but each encode the same amino acid sequence, wherein said amino acid sequences encoded by the first and second coding sequences correspond to amino acid sequences having at least 80% identity to the amino acid sequence of an endogenous human ATXN3 gene, and wherein the transgene is equal to or less than 4.7 kb.