US 2024/0181081 A1Published application

Compositions and Methods for Treatment of Myotonic Dystrophy Type 1 with CRISPR/SACAS9

AssigneeVertex Pharmaceuticals Incorporated

InventorsGuoxiang Ruan, Jianming Liu, Tudor Fulga, Eric Gunnar Anderson, Lingjun Rao, Norzehan Abdul-Manan, Matthias Heidenreich, Gregoriy Aleksandrovich Dokshin, Jesper Gromada

Life of this patent

Estimating the term…

Abstract

Compositions and methods for treating Myotonic Dystrophy Type 1 (DM1) are encompassed.

Claims38

Dependent claims

A composition comprising a single nucleic acid molecule encoding one or more guide RNAs and a Cas9, wherein the single nucleic acid molecule comprises:

a. a first nucleic acid encoding one or more spacer sequences selected from any one of SEQ ID NOs:
1-8, 10-28, and 101-154, and a second nucleic acid encoding a Staphylococcus aureus Cas9 (SaCas9);
b. a first nucleic acid encoding one or more spacer sequences comprising at least 20, or 21 contiguous nucleotides of a spacer sequence selected from any one of SEQ ID NOs:
1-8, 10-28, and 101-154, and a second nucleic acid encoding a Staphylococcus aureus Cas9 (SaCas9);
c. a first nucleic acid encoding one or more spacer sequences that are at least 90% identical to any one of SEQ ID NOs:
1-8, 10-28, and 101-154, and a second nucleic acid encoding a Staphylococcus aureus Cas9 (SaCas9);
d. a first nucleic acid encoding one or more spacer sequences selected from any one of SEQ ID NOs:
1, 2, 3, 4, 7, 8, 10, 11, 12, 13, 14, 15, 18, 19, 20, 21, 23, 25, 26, 27, and 28, and a second nucleic acid encoding a Staphylococcus aureus Cas9 (SaCas9);
e. a first nucleic acid encoding one or more spacer sequences selected from any one of SEQ ID NOs:
1, 2, 3, 4, 7, 8, 12, and 20, and a second nucleic acid encoding a Staphylococcus aureus Cas9 (SaCas9);
f. a first nucleic acid encoding one or more spacer sequences selected from any one of SEQ ID NOs:
1, 2, 3, 4, 7, 8, and 20, and a second nucleic acid encoding a Staphylococcus aureus Cas9 (SaCas9);
g. a first nucleic acid encoding one or more spacer sequences selected from any one of SEQ ID NOs:
1, 101, and 102, and a second nucleic acid encoding a Staphylococcus aureus Cas9 (SaCas9);
h. a first nucleic acid encoding a pair of guide RNAs comprising a first and second spacer sequence selected from any one of SEQ ID NOs:
1 and 10;
1 and 11;
1 and 12;
land 13;
land 14;
land 15;
land 16;
1 and 17;
1 and 18;
1 and 19;
1 and 20;
1 and 21;
1 and 22;
1 and 23;
1 and 24;
1 and 25;
1 and 26;
1 and 27;
1 and 28;
2 and 10;
2 and 11;
2 and 12;
2 and 13;
2 and 14;
2 and 15;
2 and 16;
2 and 17;
2 and 18;
2 and 19;
2 and 20;
2 and 21;
2 and 22;
2 and 23;
2 and 24;
2 and 25;
2 and 26;
2 and 27;
2 and 28;
3 and 10;
3 and 11;
3 and 12;
3 and 13;
3 and 14;
3 and 15;
3 and 16;
3 and 17;
3 and 18;
3 and 19;
3 and 20;
3 and 21;
3 and 22;
3 and 23;
3 and 24;
3 and 25;
3 and 26;
3 and 27;
3 and 28;
4 and 10;
4 and 11;
4 and 12;
4 and 13;
4 and 14;
4 and 15;
4 and 16;
4 and 17;
4 and 18;
4 and 19;
4 and 20;
4 and 21;
4 and 22;
4 and 23;
4 and 24;
4 and 25;
4 and 26;
4 and 27;
4 and 28;
5 and 10;
5 and 11;
5 and 12;
5 and 13;
5 and 14;
5 and 15;
5 and 16;
5 and 17;
5 and 18;
5 and 19;
5 and 20;
5 and 21;
5 and 22;
5 and 23;
5 and 24;
5 and 25;
5 and 26;
5 and 27;
5 and 28;
6 and 10;
6 and 11;
6 and 12;
6 and 13;
6 and 14;
6 and 15;
6 and 16;
6 and 17;
6 and 18;
6 and 19;
6 and 20;
6 and 21;
6 and 22;
6 and 23;
6 and 24;
6 and 25;
6 and 26;
6 and 27;
6 and 28;
7 and 10;
7 and 11;
7 and 12;
7 and 13;
7 and 14;
7 and 15;
7 and 16;
7 and 17;
7 and 18;
7 and 19;
7 and 20;
7 and 21;
7 and 22;
7 and 23;
7 and 24;
7 and 25;
7 and 26;
7 and 27;
7 and 28;
8 and 10;
8 and 11;
8 and 12;
8 and 13;
8 and 14;
8 and 15;
8 and 16;
8 and 17;
8 and 18;
8 and 19;
8 and 20;
8 and 21;
8 and 22;
8 and 23;
8 and 24;
8 and 25;
8 and 26;
8 and 27;
and 8 and 28, and a second nucleic acid encoding a Staphylococcus aureus Cas9 (SaCas9);
i. a first nucleic acid encoding a pair of guide RNAs comprising at least 20, or 21 contiguous nucleotides of a first and second spacer sequence selected from any one of SEQ ID NOs:
1 and 10;
1 and 11;
1 and 12;
1 and 13;
1 and 14;
1 and 15;
1 and 16;
1 and 17;
1 and 18;
1 and 19;
1 and 20;
1 and 21;
1 and 22;
1 and 23;
1 and 24;
1 and 25;
1 and 26;
1 and 27;
1 and 28;
2 and 10;
2 and 11;
2 and 12;
2 and 13;
2 and 14;
2 and 15;
2 and 16;
2 and 17;
2 and 18;
2 and 19;
2 and 20;
2 and 21;
2 and 22;
2 and 23;
2 and 24;
2 and 25;
2 and 26;
2 and 27;
2 and 28;
3 and 10;
3 and 11;
3 and 12;
3 and 13;
3 and 14;
3 and 15;
3 and 16;
3 and 17;
3 and 18;
3 and 19;
3 and 20;
3 and 21;
3 and 22;
3 and 23;
3 and 24;
3 and 25;
3 and 26;
3 and 27;
3 and 28;
4 and 10;
4 and 11;
4 and 12;
4 and 13;
4 and 14;
4 and 15;
4 and 16;
4 and 17;
4 and 18;
4 and 19;
4 and 20;
4 and 21;
4 and 22;
4 and 23;
4 and 24;
4 and 25;
4 and 26;
4 and 27;
4 and 28;
5 and 10;
5 and 11;
5 and 12;
5 and 13;
5 and 14;
5 and 15;
5 and 16;
5 and 17;
5 and 18;
5 and 19;
5 and 20;
5 and 21;
5 and 22;
5 and 23;
5 and 24;
5 and 25;
5 and 26;
5 and 27;
5 and 28;
6 and 10;
6 and 11;
6 and 12;
6 and 13;
6 and 14;
6 and 15;
6 and 16;
6 and 17;
6 and 18;
6 and 19;
6 and 20;
6 and 21;
6 and 22;
6 and 23;
6 and 24;
6 and 25;
6 and 26;
6 and 27;
6 and 28;
7 and 10;
7 and 11;
7 and 12;
7 and 13;
7 and 14;
7 and 15;
7 and 16;
7 and 17;
7 and 18;
7 and 19;
7 and 20;
7 and 21;
7 and 22;
7 and 23;
7 and 24;
7 and 25;
7 and 26;
7 and 27;
7 and 28;
8 and 10;
8 and 11;
8 and 12;
8 and 13;
8 and 14;
8 and 15;
8 and 16;
8 and 17;
8 and 18;
8 and 19;
8 and 20;
8 and 21;
8 and 22;
8 and 23;
8 and 24;
8 and 25;
8 and 26;
8 and 27;
and 8 and 28, and a second nucleic acid encoding a Staphylococcus aureus Cas9 (SaCas9);
j. a first nucleic acid encoding a pair of guide RNAs that is at least 90% identical to a first and second spacer sequence selected from any one of SEQ ID NOs:
1 and 10;
land 11;
land 12;
land 13;
land 14;
1 and 15;
1 and 16;
1 and 17;
1 and 18;
1 and 19;
1 and 20;
1 and 21;
1 and 22;
1 and 23;
1 and 24;
1 and 25;
1 and 26;
1 and 27;
1 and 28;
2 and 10;
2 and 11;
2 and 12;
2 and 13;
2 and 14;
2 and 15;
2 and 16;
2 and 17;
2 and 18;
2 and 19;
2 and 20;
2 and 21;
2 and 22;
2 and 23;
2 and 24;
2 and 25;
2 and 26;
2 and 27;
2 and 28;
3 and 10;
3 and 11;
3 and 12;
3 and 13;
3 and 14;
3 and 15;
3 and 16;
3 and 17;
3 and 18;
3 and 19;
3 and 20;
3 and 21;
3 and 22;
3 and 23;
3 and 24;
3 and 25;
3 and 26;
3 and 27;
3 and 28;
4 and 10;
4 and 11;
4 and 12;
4 and 13;
4 and 14;
4 and 15;
4 and 16;
4 and 17;
4 and 18;
4 and 19;
4 and 20;
4 and 21;
4 and 22;
4 and 23;
4 and 24;
4 and 25;
4 and 26;
4 and 27;
4 and 28;
5 and 10;
5 and 11;
5 and 12;
5 and 13;
5 and 14;
5 and 15;
5 and 16;
5 and 17;
5 and 18;
5 and 19;
5 and 20;
5 and 21;
5 and 22;
5 and 23;
5 and 24;
5 and 25;
5 and 26;
5 and 27;
5 and 28;
6 and 10;
6 and 11;
6 and 12;
6 and 13;
6 and 14;
6 and 15;
6 and 16;
6 and 17;
6 and 18;
6 and 19;
6 and 20;
6 and 21;
6 and 22;
6 and 23;
6 and 24;
6 and 25;
6 and 26;
6 and 27;
6 and 28;
7 and 10;
7 and 11;
7 and 12;
7 and 13;
7 and 14;
7 and 15;
7 and 16;
7 and 17;
7 and 18;
7 and 19;
7 and 20;
7 and 21;
7 and 22;
7 and 23;
7 and 24;
7 and 25;
7 and 26;
7 and 27;
7 and 28;
8 and 10;
8 and 11;
8 and 12;
8 and 13;
8 and 14;
8 and 15;
8 and 16;
8 and 17;
8 and 18;
8 and 19;
8 and 20;
8 and 21;
8 and 22;
8 and 23;
8 and 24;
8 and 25;
8 and 26;
8 and 27;
and 8 and 28, and a second nucleic acid encoding a Staphylococcus aureus Cas9 (SaCas9);
k. a first nucleic acid encoding a pair of guide RNAs comprising a first and second spacer sequence selected from any one of SEQ ID NOs:
4 and 12;
2 and 12;
3 and 12;
4 and 20;
4 and 18;
2 and 10;
4 and 28;
I and 12;
8 and 12;
4 and 13;
4 and 23;
3 and 10;
8 and 20;
1 and 10;
2 and 23;
2 and 20;
8 and 23;
8 and 10;
1 and 18;
2 and 13;
2 and 18;
3 and 18;
2 and 28;
7 and 12;
8 and 18;
3 and 20;
3 and 23;
2 and 13;
1 and 23;
8 and 13;
3 and 28;
8 and 28;
7 and 10;
1 and 13;
1 and 20;
1 and 28;
4 and 27;
7 and 20;
7 and 23;
7 and 13;
7 and 28;
2 and 27;
8 and 27;
4 and 11;
4 and 25;
4 and 28;
4 and 19;
4 and 15;
8 and 11;
3 and 27;
2 and 25;
2 and 11;
7 and 18;
3 and 25;
8 and 15;
8 and 25;
3 and 11;
3 and 19;
1 and 15;
3 and 15;
1 and 27;
2 and 15;
2 and 19;
1 and 11;
1 and 25;
8 and 19;
4 and 21;
8 and 21;
7 and 27;
7 and 15;
1 and 19;
2 and 21;
7 and 11;
3 and 21;
4 and 14;
7 and 19;
4 and 26;
8 and 26;
7 and 25;
1 and 21;
3 and 26;
2 and 26;
8 and 14;
1 and 14;
2 and 14;
3 and 14;
1 and 26;
7 and 21;
7 and 14;
and 7 and 26, and a second nucleic acid encoding a Staphylococcus aureus Cas9 (SaCas9);
l. a first nucleic acid encoding a pair of guide RNAs comprising a first and second spacer sequence selected from any one of SEQ ID NOs:
4 and 12;
2 and 12;
3 and 12;
4 and 20;
4 and 18;
2 and 10;
4 and 28;
I and 12;
8 and 12;
4 and 13;
4 and 23;
3 and 10;
8 and 20;
1 and 10;
2 and 23;
2 and 20;
8 and 23;
8 and 10;
1 and 18;
2 and 13;
2 and 18;
3 and 18;
2 and 28;
7 and 12;
8 and 18;
3 and 20;
3 and 23;
2 and 13;
1 and 23;
8 and 13;
3 and 28;
8 and 28;
7 and 10;
1 and 13;
1 and 20;
1 and 28;
4 and 27;
7 and 20;
7 and 23;
7 and 13;
7 and 28;
2 and 27;
8 and 12;
8 and 27;
4 and 11;
and 4 and 25, and a second nucleic acid encoding a Staphylococcus aureus Cas9 (SaCas9);
m. a first nucleic acid encoding a pair of guide RNAs comprising a first and second spacer sequence selected from any one of SEQ ID NOs:
4 and 12;
2 and 12;
3 and 12;
4 and 20;
4 and 18;
2 and 10;
4 and 28;
I and 12;
8 and 12;
4 and 13;
4 and 23;
3 and 10;
8 and 20;
1 and 10;
2 and 23;
2 and 20;
8 and 23;
1 and 18;
2 and 13;
2 and 18;
3 and 18;
2 and 28;
7 and 12;
8 and 18;
3 and 20;
3 and 23;
2 and 13;
1 and 23;
8 and 13;
3 and 28;
8 and 28;
1 and 13;
1 and 20;
1 and 28;
4 and 27;
7 and 20;
7 and 23;
7 and 13;
7 and 28;
2 and 27;
4 and 11;
4 and 25;
4 and 28;
4 and 19;
4 and 15;
8 and 11;
3 and 27;
2 and 25;
2 and 11;
7 and 18;
3 and 25;
8 and 15;
3 and 11;
3 and 19;
1 and 15;
3 and 15;
1 and 27;
2 and 15;
2 and 19;
1 and 11;
1 and 25;
4 and 21;
8 and 21;
7 and 27;
7 and 15;
1 and 19;
2 and 21;
7 and 11;
3 and 21;
7 and 19;
7 and 25;
1 and 21;
3 and 26;
3 and 14;
7 and 21;
and 7 and 14 and a second nucleic acid encoding a Staphylococcus aureus Cas9 (SaCas9);
n. a first nucleic acid encoding a pair of guide RNAs comprising a first and second spacer sequence selected from any one of SEQ ID NOs:
4 and 12;
2 and 10;
4 and 18;
4 and 28;
8 and 12;
4 and 13;
3 and 10;
7 and 12;
7 and 13;
4 and 28;
and 7 and 18, and a second nucleic acid encoding a Staphylococcus aureus Cas9 (SaCas9);
o. a first nucleic acid encoding a pair of guide RNAs comprising a first and second spacer sequence selected from any one of SEQ ID NOs:
4 and 12;
2 and 12;
3 and 12;
4 and 20;
4 and 18;
2 and 10;
4 and 10;
1 and 12;
8 and 12;
4 and 13;
7 and 12;
7 and 28;
7 and 18 and a second nucleic acid encoding a Staphylococcus aureus Cas9 (SaCas9);
p. a first nucleic acid encoding a pair of guide RNAs comprising a first and second spacer sequence of SEQ ID NOs:
7 and 12, and a second nucleic acid encoding a Staphylococcus aureus Cas9 (SaCas9);
q. a first nucleic acid encoding a pair of guide RNAs comprising a first and second spacer sequence of SEQ ID NOs:
4 and 12, and a second nucleic acid encoding a Staphylococcus aureus Cas9 (SaCas9);
or
r. a first nucleic acid encoding a pair of guide RNAs comprising a first and second spacer sequence of SEQ ID NOs:
4 and 18, and a second nucleic acid encoding a Staphylococcus aureus Cas9 (SaCas9);
or
s. a first nucleic acid encoding a pair of guide RNAs comprising a first and second spacer sequence of SEQ ID NOs:
2 and 12, and a second nucleic acid encoding a Staphylococcus aureus Cas9 (SaCas9);
or
t. a first nucleic acid encoding a pair of guide RNAs comprising a first and second spacer sequence of SEQ ID NOs:
4 and 13, and a second nucleic acid encoding a Staphylococcus aureus Cas9 (SaCas9);
or
u. a first nucleic acid encoding a pair of guide RNAs comprising a first and second spacer sequence of SEQ ID NOs:
8 and 12, and a second nucleic acid encoding a Staphylococcus aureus Cas9 (SaCas9);
or
v. a first nucleic acid encoding a pair of guide RNAs comprising a first and second spacer sequence of SEQ ID NOs:
7 and 23, and a second nucleic acid encoding a Staphylococcus aureus Cas9 (SaCas9).

730 more elements

A method of treating Myotonic Dystrophy Type 1 (DM1), the method comprising delivering to a cell a single nucleic acid molecule comprising a nucleic acid encoding a Staphylococcus aureus Cas9 (SaCas9);

and:
i) a nucleic acid encoding a guide RNA, wherein the guide RNA comprises:
a. one or more spacer sequences selected from any one of SEQ ID NOs:
1-8, 10-28, and 101-154;
b. one or more spacer sequences comprising at least 20, or 21 contiguous nucleotides of a spacer sequence selected from any one of SEQ ID NOs:
1-8, 10-28, and 101-154;
or
c. one or more spacer sequences that are at least 90% identical to any one of SEQ ID NOs:
1-8, 10-28, and 101-154;
or
ii) a nucleic acid encoding a pair of guide RNAs comprising:
a. a first and second spacer sequence selected from any one of SEQ ID NOs:
1 and 10;
land 11;
land 12;
land 13;
1 and 14;
1 and 15;
1 and 16;
1 and 17;
1 and 18;
1 and 19;
1 and 20;
1 and 21;
1 and 22;
1 and 23;
1 and 24;
1 and 25;
1 and 26;
1 and 27;
1 and 28;
2 and 10;
2 and 11;
2 and 12;
2 and 13;
2 and 14;
2 and 15;
2 and 16;
2 and 17;
2 and 18;
2 and 19;
2 and 20;
2 and 21;
2 and 22;
2 and 23;
2 and 24;
2 and 25;
2 and 26;
2 and 27;
2 and 28;
3 and 10;
3 and 11;
3 and 12;
3 and 13;
3 and 14;
3 and 15;
3 and 16;
3 and 17;
3 and 18;
3 and 19;
3 and 20;
3 and 21;
3 and 22;
3 and 23;
3 and 24;
3 and 25;
3 and 26;
3 and 27;
3 and 28;
4 and 10;
4 and 11;
4 and 12;
4 and 13;
4 and 14;
4 and 15;
4 and 16;
4 and 17;
4 and 18;
4 and 19;
4 and 20;
4 and 21;
4 and 22;
4 and 23;
4 and 24;
4 and 25;
4 and 26;
4 and 27;
4 and 28;
5 and 10;
5 and 11;
5 and 12;
5 and 13;
5 and 14;
5 and 15;
5 and 16;
5 and 17;
5 and 18;
5 and 19;
5 and 20;
5 and 21;
5 and 22;
5 and 23;
5 and 24;
5 and 25;
5 and 26;
5 and 27;
5 and 28;
6 and 10;
6 and 11;
6 and 12;
6 and 13;
6 and 14;
6 and 15;
6 and 16;
6 and 17;
6 and 18;
6 and 19;
6 and 20;
6 and 21;
6 and 22;
6 and 23;
6 and 24;
6 and 25;
6 and 26;
6 and 27;
6 and 28;
7 and 10;
7 and 11;
7 and 12;
7 and 13;
7 and 14;
7 and 15;
7 and 16;
7 and 17;
7 and 18;
7 and 19;
7 and 20;
7 and 21;
7 and 22;
7 and 23;
7 and 24;
7 and 25;
7 and 26;
7 and 27;
7 and 28;
8 and 10;
8 and 11;
8 and 12;
8 and 13;
8 and 14;
8 and 15;
8 and 16;
8 and 17;
8 and 18;
8 and 19;
8 and 20;
8 and 21;
8 and 22;
8 and 23;
8 and 24;
8 and 25;
8 and 26;
8 and 27;
and 8 and 28;
b. a first and second spacer sequence comprising at least 20, or 21 contiguous nucleotides of any of the first and second spacer sequences of i) a.;
or
c. a first and second spacer sequence that is at least 90% identical to any of the first and second spacer sequences of i) a. or i) b.

164 more elements

A method of excising a CTG repeat in the 3′ UTR of the DMPK gene, the method comprising delivering to a cell a single nucleic acid molecule comprising a nucleic acid encoding a Staphylococcus aureus Cas9 (SaCas9);

and:
i) a nucleic acid encoding a guide RNA, wherein the guide RNA comprises:
a. one or more spacer sequences selected from any one of SEQ ID NOs:
1-8, 10-28, and 101-154;
b. one or more spacer sequences comprising at least 20, or 21 contiguous nucleotides of a spacer sequence selected from any one of SEQ ID NOs:
1-8, 10-28, and 101-154;
or
c. one or more spacer sequences that are at least 90% identical to any one of SEQ ID NOs:
1-8, 10-28, and 101-154;
or
ii) a nucleic acid encoding a pair of guide RNAs comprising:
a. a first and second spacer sequence selected from SEQ ID NOs:
1 and 10;
1 and 11;
land 12;
land 13;
land 14;
1 and 15;
1 and 16;
1 and 17;
1 and 18;
1 and 19;
1 and 20;
1 and 21;
1 and 22;
1 and 23;
1 and 24;
1 and 25;
1 and 26;
1 and 27;
1 and 28;
2 and 10;
2 and 11;
2 and 12;
2 and 13;
2 and 14;
2 and 15;
2 and 16;
2 and 17;
2 and 18;
2 and 19;
2 and 20;
2 and 21;
2 and 22;
2 and 23;
2 and 24;
2 and 25;
2 and 26;
2 and 27;
2 and 28;
3 and 10;
3 and 11;
3 and 12;
3 and 13;
3 and 14;
3 and 15;
3 and 16;
3 and 17;
3 and 18;
3 and 19;
3 and 20;
3 and 21;
3 and 22;
3 and 23;
3 and 24;
3 and 25;
3 and 26;
3 and 27;
3 and 28;
4 and 10;
4 and 11;
4 and 12;
4 and 13;
4 and 14;
4 and 15;
4 and 16;
4 and 17;
4 and 18;
4 and 19;
4 and 20;
4 and 21;
4 and 22;
4 and 23;
4 and 24;
4 and 25;
4 and 26;
4 and 27;
4 and 28;
5 and 10;
5 and 11;
5 and 12;
5 and 13;
5 and 14;
5 and 15;
5 and 16;
5 and 17;
5 and 18;
5 and 19;
5 and 20;
5 and 21;
5 and 22;
5 and 23;
5 and 24;
5 and 25;
5 and 26;
5 and 27;
5 and 28;
6 and 10;
6 and 11;
6 and 12;
6 and 13;
6 and 14;
6 and 15;
6 and 16;
6 and 17;
6 and 18;
6 and 19;
6 and 20;
6 and 21;
6 and 22;
6 and 23;
6 and 24;
6 and 25;
6 and 26;
6 and 27;
6 and 28;
7 and 10;
7 and 11;
7 and 12;
7 and 13;
7 and 14;
7 and 15;
7 and 16;
7 and 17;
7 and 18;
7 and 19;
7 and 20;
7 and 21;
7 and 22;
7 and 23;
7 and 24;
7 and 25;
7 and 26;
7 and 27;
7 and 28;
8 and 10;
8 and 11;
8 and 12;
8 and 13;
8 and 14;
8 and 15;
8 and 16;
8 and 17;
8 and 18;
8 and 19;
8 and 20;
8 and 21;
8 and 22;
8 and 23;
8 and 24;
8 and 25;
8 and 26;
8 and 27;
and 8 and 28;
b. a first and second spacer sequence comprising at least 20, or 21 contiguous nucleotides of any of the first and second spacer sequences of i) a.;
or
c. a first and second spacer sequence that is at least 90% identical to any of the first and second spacer sequences of i) a. or i) b.

164 more elements

A composition comprising a first nucleic acid molecule and a second nucleic acid molecule, wherein the first nucleic acid molecule encodes a Staphylococcus aureus Cas9 (SaCas9) and the second nucleic acid molecule encodes:

one or more guide RNAs comprising:
a. a first and second spacer sequence selected from any one of SEQ ID NOs:
1 and 10;
land 11;
land 12;
land 13;
land 14;
1 and 15;
1 and 16;
1 and 17;
1 and 18;
1 and 19;
1 and 20;
1 and 21;
1 and 22;
1 and 23;
1 and 24;
1 and 25;
1 and 26;
1 and 27;
1 and 28;
2 and 10;
2 and 11;
2 and 12;
2 and 13;
2 and 14;
2 and 15;
2 and 16;
2 and 17;
2 and 18;
2 and 19;
2 and 20;
2 and 21;
2 and 22;
2 and 23;
2 and 24;
2 and 25;
2 and 26;
2 and 27;
2 and 28;
3 and 10;
3 and 11;
3 and 12;
3 and 13;
3 and 14;
3 and 15;
3 and 16;
3 and 17;
3 and 18;
3 and 19;
3 and 20;
3 and 21;
3 and 22;
3 and 23;
3 and 24;
3 and 25;
3 and 26;
3 and 27;
3 and 28;
4 and 10;
4 and 11;
4 and 12;
4 and 13;
4 and 14;
4 and 15;
4 and 16;
4 and 17;
4 and 18;
4 and 19;
4 and 20;
4 and 21;
4 and 22;
4 and 23;
4 and 24;
4 and 25;
4 and 26;
4 and 27;
4 and 28;
5 and 10;
5 and 11;
5 and 12;
5 and 13;
5 and 14;
5 and 15;
5 and 16;
5 and 17;
5 and 18;
5 and 19;
5 and 20;
5 and 21;
5 and 22;
5 and 23;
5 and 24;
5 and 25;
5 and 26;
5 and 27;
5 and 28;
6 and 10;
6 and 11;
6 and 12;
6 and 13;
6 and 14;
6 and 15;
6 and 16;
6 and 17;
6 and 18;
6 and 19;
6 and 20;
6 and 21;
6 and 22;
6 and 23;
6 and 24;
6 and 25;
6 and 26;
6 and 27;
6 and 28;
7 and 10;
7 and 11;
7 and 12;
7 and 13;
7 and 14;
7 and 15;
7 and 16;
7 and 17;
7 and 18;
7 and 19;
7 and 20;
7 and 21;
7 and 22;
7 and 23;
7 and 24;
7 and 25;
7 and 26;
7 and 27;
7 and 28;
8 and 10;
8 and 11;
8 and 12;
8 and 13;
8 and 14;
8 and 15;
8 and 16;
8 and 17;
8 and 18;
8 and 19;
8 and 20;
8 and 21;
8 and 22;
8 and 23;
8 and 24;
8 and 25;
8 and 26;
8 and 27;
and 8 and 28;
b. a first and second spacer sequence comprising at least 20, or 21 contiguous nucleotides of any of the first and second spacer sequences of a.;
c. a first and second spacer sequence that is at least 90% identical to any of the first and second spacer sequences of a. or b.

153 more elements

Dependencies are inferred from the claim text.

Facts

Application: 18/456,269Filed 25 Aug 2023
Priority: 26 Feb 2021
Published: 6 Jun 2024
Term ends: Not estimatedThe source records don’t support an estimate.

Classification15 codes

A61K 48/005
Medicinal preparations containing genetic material which is inserted into cells of the living body to treat genetic diseases; Gene therapy:…
C12N 9/12
Transferases: transferring phosphorus containing groups, e.g. kinases
C12N 9/22
Acting on ester bonds: ribonucleases [RNase]; Deoxyribonucleases [DNase]
C12N 9/226
Clustered regularly interspaced short palindromic repeats [CRISPR]-associated [CAS] enzymes: class 2 CAS enzyme complex, e.g. single CAS protein

Family

Application63/154,442Us provisional application · filed 26 Feb 2021
Application63/159,815Us provisional application · filed 11 Mar 2021
Application63/184,462Us provisional application · filed 5 May 2021
Application63/276,003Us provisional application · filed 5 Nov 2021
Application63/306,883Us provisional application · filed 4 Feb 2022
Application18/456,269Continuation
US 2024/0181081 A1This patent

From the US filing history. Foreign family members aren’t listed.

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Citations

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A composition comprising a single nucleic acid molecule encoding one or more guide RNAs and a Cas9, wherein the single nucleic acid molecule comprises:

a. a first nucleic acid encoding one or more spacer sequences selected from any one of SEQ ID NOs:
1-8, 10-28, and 101-154, and a second nucleic acid encoding a Staphylococcus aureus Cas9 (SaCas9);
b. a first nucleic acid encoding one or more spacer sequences comprising at least 20, or 21 contiguous nucleotides of a spacer sequence selected from any one of SEQ ID NOs:
1-8, 10-28, and 101-154, and a second nucleic acid encoding a Staphylococcus aureus Cas9 (SaCas9);
c. a first nucleic acid encoding one or more spacer sequences that are at least 90% identical to any one of SEQ ID NOs:
1-8, 10-28, and 101-154, and a second nucleic acid encoding a Staphylococcus aureus Cas9 (SaCas9);
d. a first nucleic acid encoding one or more spacer sequences selected from any one of SEQ ID NOs:
1, 2, 3, 4, 7, 8, 10, 11, 12, 13, 14, 15, 18, 19, 20, 21, 23, 25, 26, 27, and 28, and a second nucleic acid encoding a Staphylococcus aureus Cas9 (SaCas9);
e. a first nucleic acid encoding one or more spacer sequences selected from any one of SEQ ID NOs:
1, 2, 3, 4, 7, 8, 12, and 20, and a second nucleic acid encoding a Staphylococcus aureus Cas9 (SaCas9);
f. a first nucleic acid encoding one or more spacer sequences selected from any one of SEQ ID NOs:
1, 2, 3, 4, 7, 8, and 20, and a second nucleic acid encoding a Staphylococcus aureus Cas9 (SaCas9);
g. a first nucleic acid encoding one or more spacer sequences selected from any one of SEQ ID NOs:
1, 101, and 102, and a second nucleic acid encoding a Staphylococcus aureus Cas9 (SaCas9);
h. a first nucleic acid encoding a pair of guide RNAs comprising a first and second spacer sequence selected from any one of SEQ ID NOs:
1 and 10;
1 and 11;
1 and 12;
land 13;
land 14;
land 15;
land 16;
1 and 17;
1 and 18;
1 and 19;
1 and 20;
1 and 21;
1 and 22;
1 and 23;
1 and 24;
1 and 25;
1 and 26;
1 and 27;
1 and 28;
2 and 10;
2 and 11;
2 and 12;
2 and 13;
2 and 14;
2 and 15;
2 and 16;
2 and 17;
2 and 18;
2 and 19;
2 and 20;
2 and 21;
2 and 22;
2 and 23;
2 and 24;
2 and 25;
2 and 26;
2 and 27;
2 and 28;
3 and 10;
3 and 11;
3 and 12;
3 and 13;
3 and 14;
3 and 15;
3 and 16;
3 and 17;
3 and 18;
3 and 19;
3 and 20;
3 and 21;
3 and 22;
3 and 23;
3 and 24;
3 and 25;
3 and 26;
3 and 27;
3 and 28;
4 and 10;
4 and 11;
4 and 12;
4 and 13;
4 and 14;
4 and 15;
4 and 16;
4 and 17;
4 and 18;
4 and 19;
4 and 20;
4 and 21;
4 and 22;
4 and 23;
4 and 24;
4 and 25;
4 and 26;
4 and 27;
4 and 28;
5 and 10;
5 and 11;
5 and 12;
5 and 13;
5 and 14;
5 and 15;
5 and 16;
5 and 17;
5 and 18;
5 and 19;
5 and 20;
5 and 21;
5 and 22;
5 and 23;
5 and 24;
5 and 25;
5 and 26;
5 and 27;
5 and 28;
6 and 10;
6 and 11;
6 and 12;
6 and 13;
6 and 14;
6 and 15;
6 and 16;
6 and 17;
6 and 18;
6 and 19;
6 and 20;
6 and 21;
6 and 22;
6 and 23;
6 and 24;
6 and 25;
6 and 26;
6 and 27;
6 and 28;
7 and 10;
7 and 11;
7 and 12;
7 and 13;
7 and 14;
7 and 15;
7 and 16;
7 and 17;
7 and 18;
7 and 19;
7 and 20;
7 and 21;
7 and 22;
7 and 23;
7 and 24;
7 and 25;
7 and 26;
7 and 27;
7 and 28;
8 and 10;
8 and 11;
8 and 12;
8 and 13;
8 and 14;
8 and 15;
8 and 16;
8 and 17;
8 and 18;
8 and 19;
8 and 20;
8 and 21;
8 and 22;
8 and 23;
8 and 24;
8 and 25;
8 and 26;
8 and 27;
and 8 and 28, and a second nucleic acid encoding a Staphylococcus aureus Cas9 (SaCas9);
i. a first nucleic acid encoding a pair of guide RNAs comprising at least 20, or 21 contiguous nucleotides of a first and second spacer sequence selected from any one of SEQ ID NOs:
1 and 10;
1 and 11;
1 and 12;
1 and 13;
1 and 14;
1 and 15;
1 and 16;
1 and 17;
1 and 18;
1 and 19;
1 and 20;
1 and 21;
1 and 22;
1 and 23;
1 and 24;
1 and 25;
1 and 26;
1 and 27;
1 and 28;
2 and 10;
2 and 11;
2 and 12;
2 and 13;
2 and 14;
2 and 15;
2 and 16;
2 and 17;
2 and 18;
2 and 19;
2 and 20;
2 and 21;
2 and 22;
2 and 23;
2 and 24;
2 and 25;
2 and 26;
2 and 27;
2 and 28;
3 and 10;
3 and 11;
3 and 12;
3 and 13;
3 and 14;
3 and 15;
3 and 16;
3 and 17;
3 and 18;
3 and 19;
3 and 20;
3 and 21;
3 and 22;
3 and 23;
3 and 24;
3 and 25;
3 and 26;
3 and 27;
3 and 28;
4 and 10;
4 and 11;
4 and 12;
4 and 13;
4 and 14;
4 and 15;
4 and 16;
4 and 17;
4 and 18;
4 and 19;
4 and 20;
4 and 21;
4 and 22;
4 and 23;
4 and 24;
4 and 25;
4 and 26;
4 and 27;
4 and 28;
5 and 10;
5 and 11;
5 and 12;
5 and 13;
5 and 14;
5 and 15;
5 and 16;
5 and 17;
5 and 18;
5 and 19;
5 and 20;
5 and 21;
5 and 22;
5 and 23;
5 and 24;
5 and 25;
5 and 26;
5 and 27;
5 and 28;
6 and 10;
6 and 11;
6 and 12;
6 and 13;
6 and 14;
6 and 15;
6 and 16;
6 and 17;
6 and 18;
6 and 19;
6 and 20;
6 and 21;
6 and 22;
6 and 23;
6 and 24;
6 and 25;
6 and 26;
6 and 27;
6 and 28;
7 and 10;
7 and 11;
7 and 12;
7 and 13;
7 and 14;
7 and 15;
7 and 16;
7 and 17;
7 and 18;
7 and 19;
7 and 20;
7 and 21;
7 and 22;
7 and 23;
7 and 24;
7 and 25;
7 and 26;
7 and 27;
7 and 28;
8 and 10;
8 and 11;
8 and 12;
8 and 13;
8 and 14;
8 and 15;
8 and 16;
8 and 17;
8 and 18;
8 and 19;
8 and 20;
8 and 21;
8 and 22;
8 and 23;
8 and 24;
8 and 25;
8 and 26;
8 and 27;
and 8 and 28, and a second nucleic acid encoding a Staphylococcus aureus Cas9 (SaCas9);
j. a first nucleic acid encoding a pair of guide RNAs that is at least 90% identical to a first and second spacer sequence selected from any one of SEQ ID NOs:
1 and 10;
land 11;
land 12;
land 13;
land 14;
1 and 15;
1 and 16;
1 and 17;
1 and 18;
1 and 19;
1 and 20;
1 and 21;
1 and 22;
1 and 23;
1 and 24;
1 and 25;
1 and 26;
1 and 27;
1 and 28;
2 and 10;
2 and 11;
2 and 12;
2 and 13;
2 and 14;
2 and 15;
2 and 16;
2 and 17;
2 and 18;
2 and 19;
2 and 20;
2 and 21;
2 and 22;
2 and 23;
2 and 24;
2 and 25;
2 and 26;
2 and 27;
2 and 28;
3 and 10;
3 and 11;
3 and 12;
3 and 13;
3 and 14;
3 and 15;
3 and 16;
3 and 17;
3 and 18;
3 and 19;
3 and 20;
3 and 21;
3 and 22;
3 and 23;
3 and 24;
3 and 25;
3 and 26;
3 and 27;
3 and 28;
4 and 10;
4 and 11;
4 and 12;
4 and 13;
4 and 14;
4 and 15;
4 and 16;
4 and 17;
4 and 18;
4 and 19;
4 and 20;
4 and 21;
4 and 22;
4 and 23;
4 and 24;
4 and 25;
4 and 26;
4 and 27;
4 and 28;
5 and 10;
5 and 11;
5 and 12;
5 and 13;
5 and 14;
5 and 15;
5 and 16;
5 and 17;
5 and 18;
5 and 19;
5 and 20;
5 and 21;
5 and 22;
5 and 23;
5 and 24;
5 and 25;
5 and 26;
5 and 27;
5 and 28;
6 and 10;
6 and 11;
6 and 12;
6 and 13;
6 and 14;
6 and 15;
6 and 16;
6 and 17;
6 and 18;
6 and 19;
6 and 20;
6 and 21;
6 and 22;
6 and 23;
6 and 24;
6 and 25;
6 and 26;
6 and 27;
6 and 28;
7 and 10;
7 and 11;
7 and 12;
7 and 13;
7 and 14;
7 and 15;
7 and 16;
7 and 17;
7 and 18;
7 and 19;
7 and 20;
7 and 21;
7 and 22;
7 and 23;
7 and 24;
7 and 25;
7 and 26;
7 and 27;
7 and 28;
8 and 10;
8 and 11;
8 and 12;
8 and 13;
8 and 14;
8 and 15;
8 and 16;
8 and 17;
8 and 18;
8 and 19;
8 and 20;
8 and 21;
8 and 22;
8 and 23;
8 and 24;
8 and 25;
8 and 26;
8 and 27;
and 8 and 28, and a second nucleic acid encoding a Staphylococcus aureus Cas9 (SaCas9);
k. a first nucleic acid encoding a pair of guide RNAs comprising a first and second spacer sequence selected from any one of SEQ ID NOs:
4 and 12;
2 and 12;
3 and 12;
4 and 20;
4 and 18;
2 and 10;
4 and 28;
I and 12;
8 and 12;
4 and 13;
4 and 23;
3 and 10;
8 and 20;
1 and 10;
2 and 23;
2 and 20;
8 and 23;
8 and 10;
1 and 18;
2 and 13;
2 and 18;
3 and 18;
2 and 28;
7 and 12;
8 and 18;
3 and 20;
3 and 23;
2 and 13;
1 and 23;
8 and 13;
3 and 28;
8 and 28;
7 and 10;
1 and 13;
1 and 20;
1 and 28;
4 and 27;
7 and 20;
7 and 23;
7 and 13;
7 and 28;
2 and 27;
8 and 27;
4 and 11;
4 and 25;
4 and 28;
4 and 19;
4 and 15;
8 and 11;
3 and 27;
2 and 25;
2 and 11;
7 and 18;
3 and 25;
8 and 15;
8 and 25;
3 and 11;
3 and 19;
1 and 15;
3 and 15;
1 and 27;
2 and 15;
2 and 19;
1 and 11;
1 and 25;
8 and 19;
4 and 21;
8 and 21;
7 and 27;
7 and 15;
1 and 19;
2 and 21;
7 and 11;
3 and 21;
4 and 14;
7 and 19;
4 and 26;
8 and 26;
7 and 25;
1 and 21;
3 and 26;
2 and 26;
8 and 14;
1 and 14;
2 and 14;
3 and 14;
1 and 26;
7 and 21;
7 and 14;
and 7 and 26, and a second nucleic acid encoding a Staphylococcus aureus Cas9 (SaCas9);
l. a first nucleic acid encoding a pair of guide RNAs comprising a first and second spacer sequence selected from any one of SEQ ID NOs:
4 and 12;
2 and 12;
3 and 12;
4 and 20;
4 and 18;
2 and 10;
4 and 28;
I and 12;
8 and 12;
4 and 13;
4 and 23;
3 and 10;
8 and 20;
1 and 10;
2 and 23;
2 and 20;
8 and 23;
8 and 10;
1 and 18;
2 and 13;
2 and 18;
3 and 18;
2 and 28;
7 and 12;
8 and 18;
3 and 20;
3 and 23;
2 and 13;
1 and 23;
8 and 13;
3 and 28;
8 and 28;
7 and 10;
1 and 13;
1 and 20;
1 and 28;
4 and 27;
7 and 20;
7 and 23;
7 and 13;
7 and 28;
2 and 27;
8 and 12;
8 and 27;
4 and 11;
and 4 and 25, and a second nucleic acid encoding a Staphylococcus aureus Cas9 (SaCas9);
m. a first nucleic acid encoding a pair of guide RNAs comprising a first and second spacer sequence selected from any one of SEQ ID NOs:
4 and 12;
2 and 12;
3 and 12;
4 and 20;
4 and 18;
2 and 10;
4 and 28;
I and 12;
8 and 12;
4 and 13;
4 and 23;
3 and 10;
8 and 20;
1 and 10;
2 and 23;
2 and 20;
8 and 23;
1 and 18;
2 and 13;
2 and 18;
3 and 18;
2 and 28;
7 and 12;
8 and 18;
3 and 20;
3 and 23;
2 and 13;
1 and 23;
8 and 13;
3 and 28;
8 and 28;
1 and 13;
1 and 20;
1 and 28;
4 and 27;
7 and 20;
7 and 23;
7 and 13;
7 and 28;
2 and 27;
4 and 11;
4 and 25;
4 and 28;
4 and 19;
4 and 15;
8 and 11;
3 and 27;
2 and 25;
2 and 11;
7 and 18;
3 and 25;
8 and 15;
3 and 11;
3 and 19;
1 and 15;
3 and 15;
1 and 27;
2 and 15;
2 and 19;
1 and 11;
1 and 25;
4 and 21;
8 and 21;
7 and 27;
7 and 15;
1 and 19;
2 and 21;
7 and 11;
3 and 21;
7 and 19;
7 and 25;
1 and 21;
3 and 26;
3 and 14;
7 and 21;
and 7 and 14 and a second nucleic acid encoding a Staphylococcus aureus Cas9 (SaCas9);
n. a first nucleic acid encoding a pair of guide RNAs comprising a first and second spacer sequence selected from any one of SEQ ID NOs:
4 and 12;
2 and 10;
4 and 18;
4 and 28;
8 and 12;
4 and 13;
3 and 10;
7 and 12;
7 and 13;
4 and 28;
and 7 and 18, and a second nucleic acid encoding a Staphylococcus aureus Cas9 (SaCas9);
o. a first nucleic acid encoding a pair of guide RNAs comprising a first and second spacer sequence selected from any one of SEQ ID NOs:
4 and 12;
2 and 12;
3 and 12;
4 and 20;
4 and 18;
2 and 10;
4 and 10;
1 and 12;
8 and 12;
4 and 13;
7 and 12;
7 and 28;
7 and 18 and a second nucleic acid encoding a Staphylococcus aureus Cas9 (SaCas9);
p. a first nucleic acid encoding a pair of guide RNAs comprising a first and second spacer sequence of SEQ ID NOs:
7 and 12, and a second nucleic acid encoding a Staphylococcus aureus Cas9 (SaCas9);
q. a first nucleic acid encoding a pair of guide RNAs comprising a first and second spacer sequence of SEQ ID NOs:
4 and 12, and a second nucleic acid encoding a Staphylococcus aureus Cas9 (SaCas9);
or
r. a first nucleic acid encoding a pair of guide RNAs comprising a first and second spacer sequence of SEQ ID NOs:
4 and 18, and a second nucleic acid encoding a Staphylococcus aureus Cas9 (SaCas9);
or
s. a first nucleic acid encoding a pair of guide RNAs comprising a first and second spacer sequence of SEQ ID NOs:
2 and 12, and a second nucleic acid encoding a Staphylococcus aureus Cas9 (SaCas9);
or
t. a first nucleic acid encoding a pair of guide RNAs comprising a first and second spacer sequence of SEQ ID NOs:
4 and 13, and a second nucleic acid encoding a Staphylococcus aureus Cas9 (SaCas9);
or
u. a first nucleic acid encoding a pair of guide RNAs comprising a first and second spacer sequence of SEQ ID NOs:
8 and 12, and a second nucleic acid encoding a Staphylococcus aureus Cas9 (SaCas9);
or
v. a first nucleic acid encoding a pair of guide RNAs comprising a first and second spacer sequence of SEQ ID NOs:
7 and 23, and a second nucleic acid encoding a Staphylococcus aureus Cas9 (SaCas9).

730 more elements

Facts

Application

18/456,269Filed 25 Aug 2023

Priority

26 Feb 2021

Published

6 Jun 2024

Term ends

Not estimatedThe source records don’t support an estimate.

Classification15 codes

A61K 48/005
Medicinal preparations containing genetic material which is inserted into cells of the living body to treat genetic diseases; Gene therapy:…
C12N 9/12
Transferases: transferring phosphorus containing groups, e.g. kinases
C12N 9/22
Acting on ester bonds: ribonucleases [RNase]; Deoxyribonucleases [DNase]
C12N 9/226
Clustered regularly interspaced short palindromic repeats [CRISPR]-associated [CAS] enzymes: class 2 CAS enzyme complex, e.g. single CAS protein

Family

Application63/154,442Us provisional application · filed 26 Feb 2021

Application63/159,815Us provisional application · filed 11 Mar 2021

Application63/184,462Us provisional application · filed 5 May 2021

Application63/276,003Us provisional application · filed 5 Nov 2021

Application63/306,883Us provisional application · filed 4 Feb 2022

Application18/456,269Continuation

US 2024/0181081 A1This patent

From the US filing history. Foreign family members aren’t listed.