US 12,214,054 B2
Therapeutic targets for the correction of the human dystrophin gene by gene editing and methods of use
Charles A. Gersbach, Durham, NC (US); and Jacqueline N. Robinson-Hamm, Durham, NC (US)
Assigned to Duke University, Durham, NC (US)
Appl. No. 15/779,633
Filed by Duke University, Durham, NC (US)
PCT Filed Nov. 30, 2016, PCT No. PCT/US2016/064285
§ 371(c)(1), (2) Date May 29, 2018,
PCT Pub. No. WO2017/095967, PCT Pub. Date Jun. 8, 2017.
Claims priority of provisional application 62/330,336, filed on May 2, 2016.
Claims priority of provisional application 62/260,712, filed on Nov. 30, 2015.
Prior Publication US 2018/0353615 A1, Dec. 13, 2018
Int. Cl. A61K 48/00 (2006.01); A61K 31/7105 (2006.01); A61K 47/26 (2006.01); A61K 47/54 (2017.01); C12N 15/10 (2006.01); C12N 15/113 (2010.01); C12N 15/80 (2006.01); C12N 15/85 (2006.01); C12Q 1/68 (2018.01)
CPC A61K 48/005 (2013.01) [A61K 31/7105 (2013.01); A61K 47/26 (2013.01); A61K 47/549 (2017.08); A61K 48/0016 (2013.01); C12N 15/102 (2013.01); C12N 15/113 (2013.01); C12N 15/80 (2013.01); C12N 15/8509 (2013.01); C12Q 1/68 (2013.01); C12N 2310/20 (2017.05)] 16 Claims
 
1. A DNA targeting composition comprising a first gRNA molecule and a second gRNA molecule, wherein the first gRNA molecule and the second gRNA molecule are selected from the group consisting of:
(i) a first gRNA molecule comprising a targeting domain that is encoded by a polynucleotide comprising a nucleotide sequence set forth in SEQ ID NO: 1, and a second gRNA molecule comprising a targeting domain that is encoded by a polynucleotide comprising a nucleotide sequence set forth in SEQ ID NO: 2;
(ii) a first gRNA molecule comprising a targeting domain that is encoded by a polynucleotide comprising a nucleotide sequence set forth in SEQ ID NO: 11, and a second gRNA molecule comprising a targeting domain that is encoded by a polynucleotide comprising a nucleotide sequence set forth in SEQ ID NO: 4;
(iii) a first gRNA molecule comprising a targeting domain that is encoded by a polynucleotide comprising a nucleotide sequence set forth in SEQ ID NO: 15, and a second gRNA molecule comprising a targeting domain that is encoded by a polynucleotide comprising a nucleotide sequence set forth in SEQ ID NO: 19;
(iv) a first gRNA molecule comprising a targeting domain that is encoded by a polynucleotide comprising a nucleotide sequence set forth in SEQ ID NO: 15, and a second gRNA molecule comprising a targeting domain that is encoded by a polynucleotide comprising a nucleotide sequence set forth in SEQ ID NO: 18;
(v) a first gRNA molecule comprising a targeting domain that is encoded by a polynucleotide comprising a nucleotide sequence set forth in SEQ ID NO: 15, and a second gRNA molecule comprising a targeting domain that is encoded by a polynucleotide comprising a nucleotide sequence set forth in SEQ ID NO: 4;
(vi) a first gRNA molecule comprising a targeting domain that is encoded by a polynucleotide comprising a nucleotide sequence set forth in SEQ ID NO: 14, and a second gRNA molecule comprising a targeting domain that is encoded by a polynucleotide comprising a nucleotide sequence set forth in SEQ ID NO: 19;
(vii) a first gRNA molecule comprising a targeting domain that is encoded by a polynucleotide comprising a nucleotide sequence set forth in SEQ ID NO: 14, and a second gRNA molecule comprising a targeting domain that is encoded by a polynucleotide comprising a nucleotide sequence set forth in SEQ ID NO: 18;
(viii) a first gRNA molecule comprising a targeting domain that is encoded by a polynucleotide comprising a nucleotide sequence set forth in SEQ ID NO: 14, and a second gRNA molecule comprising a targeting domain that is encoded by a polynucleotide comprising a nucleotide sequence set forth in SEQ ID NO: 4;
(ix) a first gRNA molecule comprising a targeting domain that is encoded by a polynucleotide comprising a nucleotide sequence set forth in SEQ ID NO: 11, and a second gRNA molecule comprising a targeting domain that is encoded by a polynucleotide comprising a nucleotide sequence set forth in SEQ ID NO: 19;
(x) a first gRNA molecule comprising a targeting domain that is encoded by a polynucleotide comprising a nucleotide sequence set forth in SEQ ID NO: 14, and a second gRNA molecule comprising a targeting domain that is encoded by a polynucleotide comprising a nucleotide sequence set forth in SEQ ID NO: 15;
(xi) a first gRNA molecule comprising a targeting domain that is encoded by a polynucleotide comprising a nucleotide sequence set forth in SEQ ID NO: 11, and a second gRNA molecule comprising a targeting domain that is encoded by a polynucleotide comprising a nucleotide sequence set forth in SEQ ID NO: 18; and
(xii) a first gRNA molecule comprising a targeting domain that is encoded by a polynucleotide comprising a nucleotide sequence set forth in SEQ ID NO: 41, and a second gRNA molecule comprising a targeting domain that is encoded by a polynucleotide comprising a nucleotide sequence set forth in SEQ ID NO: 42.