US 12,247,201 B2
Materials and methods for treatment of autosomal dominant retinitis pigmentosa
Albena Kantardzhieva, Cambridge, MA (US); Akiko Noma, Cambridge, MA (US); Abraham Scaria, Cambridge, MA (US); and Ryo Takeuchi, Cambridge, MA (US)
Assigned to CRISPR THERAPEUTICS AG, Zug (CH); and Bayer Healthcare LLC, Whippany, NJ (US)
Filed by CRISPR THERAPEUTICS AG, Zug (CH); and Bayer Healthcare LLC, Whippany, NJ (US)
Filed on Apr. 2, 2020, as Appl. No. 16/838,545.
Application 16/838,545 is a continuation of application No. 16/198,361, filed on Nov. 21, 2018, granted, now 10,662,425.
Claims priority of provisional application 62/724,319, filed on Aug. 29, 2018.
Claims priority of provisional application 62/693,080, filed on Jul. 2, 2018.
Claims priority of provisional application 62/649,133, filed on Mar. 28, 2018.
Claims priority of provisional application 62/589,111, filed on Nov. 21, 2017.
Prior Publication US 2020/0332288 A1, Oct. 22, 2020
This patent is subject to a terminal disclaimer.
Int. Cl. A61P 27/02 (2006.01); A61K 31/7088 (2006.01); A61K 38/46 (2006.01); C12N 9/22 (2006.01); C12N 15/11 (2006.01); C12N 15/113 (2010.01); C12N 15/90 (2006.01)
CPC C12N 15/11 (2013.01) [A61K 31/7088 (2013.01); A61K 38/465 (2013.01); A61P 27/02 (2018.01); C12N 9/22 (2013.01); C12N 15/1138 (2013.01); C12N 15/907 (2013.01); C12N 2310/20 (2017.05); C12N 2320/34 (2013.01); C12N 2800/80 (2013.01)] 30 Claims
 
1. A method for editing a P23H mutation within a RHO gene of a cell, the method comprising, contacting the cell with a self-inactivating CRISPR-Cas system comprising:
(a) a first segment comprising a nucleotide sequence that encodes an S. aureus Cas9 polypeptide;
(b) a second segment comprising a nucleotide sequence that encodes a guide RNA (gRNA) or a single-molecule guide RNA (sgRNA), wherein the gRNA or the sgRNA comprises SEQ ID NO: 5290 or 5291; and
(c) one or more third segments comprising a self-inactivating (SIN) site, wherein the SIN site is:
(i) a 5′ SIN site located upstream of the nucleotide sequence the encodes the Cas9 polypeptide and downstream of a SV40 nuclear localization signal (NLS);
(ii) a 5′ SIN site located upstream of the nucleotide sequence the encodes the Cas9 polypeptide and upstream of a SV40 NLS within a 5′UTR; or
(iii) a 3′ SIN site located within a naturally occurring or chimeric inserted intron located within the nucleotide sequence the encodes the Cas9 polypeptide,
wherein the gRNA or sgRNA is complementary to the SIN site.