US 12,416,018 B2
Regulation of gene expression by aptamer-mediated modulation of alternative splicing
Alex R. Boyne, Jersey City, NJ (US); Olivier F. Danos, New York, NY (US); Michael J. Volles, Cambridge, MA (US); and Xuecui Guo, Oyster Bay, NY (US)
Assigned to MEIRAGTX GENE REGULATION LIMITED, London (GB)
Filed by MEIRAGTX GENE REGULATION LIMITED, London (GB)
Filed on Feb. 14, 2022, as Appl. No. 17/671,048.
Application 16/692,928 is a division of application No. 15/548,043, granted, now 10,494,646, issued on Dec. 3, 2019, previously published as PCT/US2016/016234, filed on Feb. 2, 2016.
Application 17/671,048 is a continuation of application No. 16/692,928, filed on Nov. 22, 2019, granted, now 11,248,239, issued on Feb. 15, 2022.
Claims priority of provisional application 62/110,919, filed on Feb. 2, 2015.
Prior Publication US 2022/0282278 A1, Sep. 8, 2022
This patent is subject to a terminal disclaimer.
Int. Cl. C12N 15/113 (2010.01); A61K 9/00 (2006.01); A61K 38/44 (2006.01); A61K 48/00 (2006.01); C12N 9/06 (2006.01); C12N 15/115 (2010.01); C12N 15/63 (2006.01); C12N 15/86 (2006.01)
CPC C12N 15/86 (2013.01) [A61K 9/0048 (2013.01); A61K 38/44 (2013.01); A61K 48/0033 (2013.01); A61K 48/0066 (2013.01); A61K 48/0075 (2013.01); C12N 9/003 (2013.01); C12N 15/115 (2013.01); C12N 15/63 (2013.01); C12Y 105/01003 (2013.01); C12N 2310/16 (2013.01); C12N 2310/3519 (2013.01); C12N 2320/32 (2013.01); C12N 2320/33 (2013.01); C12N 2750/14143 (2013.01); C12N 2750/14171 (2013.01); C12N 2840/002 (2013.01); C12N 2840/44 (2013.01)] 43 Claims
 
1. A polynucleotide cassette for the regulation of the expression of a target gene comprising:
a. a riboswitch;
b. an alternatively-spliced exon, flanked by a 5′ intron and a 3′ intron,
wherein the riboswitch comprises (i) an effector region comprising a stem that includes the 5′ splice site sequence of the 3′ intron, and (ii) an aptamer;
wherein the 5′ splice site sequence of the 3′ intron comprises the sequence (i) GTAATG, (ii) GTRAGT, wherein R can be A or G, (iii) GTAAGC, or (iv) GTGTGG; and
wherein the alternatively-spliced exon comprises a stop codon that is in-frame with the target gene when the alternatively-spliced exon is spliced into the target gene mRNA.