US 12,215,316 B2
Materials and methods for treatment of usher syndrome type 2A
Albena Kantardzhieva, Cambridge, MA (US); Akiko Noma, Cambridge, MA (US); and Abraham Scaria, 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 Jun. 28, 2021, as Appl. No. 17/360,578.
Application 17/360,578 is a continuation of application No. 16/905,145, filed on Jun. 18, 2020, granted, now 11,072,792.
Application 16/905,145 is a continuation of application No. PCT/IB2018/060546, filed on Dec. 21, 2018.
Claims priority of provisional application 62/746,226, filed on Oct. 16, 2018.
Claims priority of provisional application 62/609,333, filed on Dec. 21, 2017.
Prior Publication US 2021/0403905 A1, Dec. 30, 2021
This patent is subject to a terminal disclaimer.
Int. Cl. C12N 15/11 (2006.01); A61P 27/00 (2006.01); C12N 9/22 (2006.01)
CPC C12N 15/11 (2013.01) [A61P 27/00 (2018.01); C12N 9/22 (2013.01); C12N 2310/20 (2017.05); C12N 2800/80 (2013.01)] 7 Claims
 
1. An in vivo method for treating a patient with Usher Syndrome Type 2A, the method comprising: editing an USH2A gene containing an IVS40 mutation in eye cells of the patient, wherein the editing comprises introducing into the eye cell one or more DNA endonucleases selected from the group consisting of Cas9 and Cpf1 endonucleases and nickases and one or more guide ribonucleic acids (gRNAs) comprising a spacer sequence complementary to a sequence with or near intron 40 of the USH2A gene to effect one or more SSBs or DSBs within or near intron 40 or the USH2A gene the results in a correction and results in restoration of usherin protein function.
 
2. A gRNA or sgRNA comprising a nucleotide sequence set forth in SEQ ID NO: 5321, SEQ ID NO: 5323, SEQ ID NO: 5325, SEQ ID NO: 5327, or SEQ ID NO: 5328.
 
3. A gRNA or sgRNA comprising;
a) SEQ ID NO: 5321 and any one of SEQ ID NOs: 5267-5269;
b) SEQ ID NO: 5323 and any one of SEQ ID NOs: 5267-5269;
c) SEQ ID NO: 5325 and any one of SEQ ID NOs: 5267-5269;
d) SEQ ID NO: 5327 and any one of SEQ ID NOs: 5267-5269; or
e) SEQ ID NO: 5328 and any one of SEQ ID NOs: 5267-5269.
 
4. A method for treating a patient with an USH2A gene containing an IVS40 mutation, the method comprising: administering an endonuclease selected from the group consisting of a Cas9 endonuclease and nickases and a Cfp1 endonuclease and a gRNA or sgRNA to eye cells of the patient, wherein the gRNA or sgRNA comprises SEQ ID NO: 5321, SEQ ID NO: 5323, SEQ ID NO: 5325, SEQ ID NO: 5327, or SEQ ID NO: 5328.
 
5. A method for treating a patient with an USH2A gene containing an IVS40 mutation, the method comprising: administering an endonuclease selected from the group consisting of a Cas9 endonuclease and nickases and a Cfp1 endonuclease and a gRNA or sgRNA to eye cells of the patient, wherein the gRNA or sgRNA comprises
a) SEQ ID NO: 5321 and any one of SEQ ID NOs: 5267-5269;
b) SEQ ID NO: 5323 and any one of SEQ ID NOs: 5267-5269;
c) SEQ ID NO: 5325 and any one of SEQ ID NOs: 5267-5269;
d) SEQ ID NO: 5327 and any one of SEQ ID NOs: 5267-5269; or
e) SEQ ID NO: 5328 and any one of SEQ ID NOs: 5267-5269.