Granted Patent B2

US 12,590,129 · App. 17/295,208 · Granted Mar 31, 2026

Inventors: Michael Roberts (Midlothian, GB); Juan Manuel Iglesias (Midlothian, GB); Jorge Omar Yanez-Cuna (Midlothian, GB); Nicolle Kippen (Midlothian, GB); Ross Fraser (Midlothian, GB); Katie Baker (Midlothian, GB)

C07K14/4705A61K48/0058C12N5/067C12N15/86C12N2830/008

Abstract

Claims (98)

1 . A synthetic liver-specific cis-regulatory module (CRM) comprising two or more operably linked cis-regulatory elements (CREs) selected from the group consisting of:

SEQ ID NO: 1;

SEQ ID NO: 2;

SEQ ID NO: 3;

SEQ ID NO: 4;

SEQ ID NO: 5;

SEQ ID NO: 7;

SEQ ID NO: 10; and

SEQ ID NO: 11.

2 . The synthetic liver-specific CRM of claim 1 comprising three or more, four or more, five or more, or six or more of said CREs.

3 . The synthetic liver-specific CRM of claim 1 comprising a combination of CREs selected from the group consisting of:

CRE0051 and CRE0058;

CRE0051 and CRE0042;

CRE0051, CRE0058 and CRE0065;

CRE0051, CRE0058 and CRE0066;

CRE0051, CRE0058, CRE0065 and CRE0066;

CRE0018, CRE0051, CRE0058, CRE0065 and CRE0066

CRE0051, CRE0065 and CRE0066;

CRE0051, CRE0074 and CRE0058;

CRE0051, CRE0074, CRE0058 and CRE0065;

CRE0051, CRE0074, CRE0058, CRE0065 and CRE0066;

CRE0058 and CRE0065;

CRE0068 and CRE0042;

CRE0058, CRE0065 and CRE0066;

CRE0074, CRE0058 and CRE0065; and

CRE0074, CRE0058, CRE0065 and CRE0066, wherein the CREs are present in the CRM in the recited order, and wherein they are adjacent to one another.

4 . The synthetic liver-specific CRM of claim 1 comprising two, three, four or more CREs selected from the group consisting of:

CRE0051;

CRE0058;

CRE0065;

CRE0066; and

CRE0074.

5 . The synthetic liver-specific CRM of claim 1 further comprising one or more CREs selected from the group consisting of:

SEQ ID NO: 12;

SEQ ID NO: 13;

SEQ ID NO: 14;

SEQ ID NO: 15;

SEQ ID NO: 16;

SEQ ID NO: 17;

SEQ ID NO: 18;

SEQ ID NO: 19;

SEQ ID NO: 20;

SEQ ID NO: 21; and

SEQ ID NO: 22.

6 . The synthetic liver-specific CRM of claim 1 comprising a combination of CREs selected from the group consisting of:

CRE0018, CRE0077, CRE0074, CRE0058, CRE0065;

CRE0068, CRE0042;

CRE0051, CRE0058;

CRE0051, CRE0042;

CRE0065, CRE0051, CRE0083.1;

CRE0018, CRE0051, CRE0058, CRE0065, CRE0066;

CRE0012, CRE0051, CRE0058, CRE0065, CRE0066;

CRE0051, CRE0058, CRE0065, CRE0066;

CRE0051, CRE0058, CRE0018;

CRE0051, CRE0058, CRE0065, CRE0018;

CRE0051, CRE0058, CRE0065, CRE0012;

CRE0047, CRE0051, CRE0058, CRE0065, CRE0066;

CRE0051, CRE0074, CRE0058, CRE0065, CRE0066;

CRE0051, CRE0058, CRE0065, CRE0001;

CRE0051, CRE0058, CRE0065; and

CRE0051, CRE0066.2, wherein the CREs are present in the CRM in the recited order, and wherein they are adjacent to one another.

7 . The synthetic liver-specific CRM of claim 1 comprising a combination of CREs selected from the group consisting of: CRE0051, CRE0066.2; CRE0065, CRE0051, CRE0083.1; CRE0065, CRE0066.2; CRE0066, CRE0066; CRE0065, CRE0066; CRE0074, CRE0058, CRE0065.1; CRE0051, CRE0074, CRE0058, CRE0065.1; CRE0077, CRE0074, CRE0058, CRE0065.1; CRE0078, CRE0074, CRE0058, CRE0065.1; CRE0074, CRE0074, CRE0065; CRE0058, CRE0065, CRE0066; CRE0074, CRE0058, CRE0065, CRE0066; CRE0074, CRE0058; CRE0018, CRE0051, CRE0058, CRE0065, CRE0066; CRE0051, CRE0058, CRE0065, CRE0066; CRE0051, CRE0058, CRE0065; CRE0012, CRE0051, CRE0058, CRE0065, CRE0066; CRE0051, CRE0058, CRE0065, CRE0012; CRE0051, CRE0058, CRE0065, CRE0018; CRE0051, CRE0058, CRE0065, CRE0001; CRE0051, CRE0058, CRE0065, CRE0077; CRE0051, CRE0058, CRE0018; CRE0047, CRE0051, CRE0058, CRE0065, CRE0066; CRE0051, CRE0058, CRE0065, CRE0066; CRE0077, CRE0058, CRE0065, CRE0066; CRE0078, CRE0058, CRE0065, CRE0066; CRE0051, CRE0074, CRE0058, CRE0065, CRE0066; CRE0077, CRE0074, CRE0058, CRE0065, CRE0066; CRE0078, CRE0074, CRE0058, CRE0065, CRE0066; CRE0051, CRE0074, CRE0058; CRE0077, CRE0074, CRE0058; CRE0078, CRE0074, CRE0058; CRE0051, CRE0058; CRE0068, CRE0042; CRE0051, CRE0058, CRE0066; CRE0051, CRE0065, CRE0066; CRE0018, CRE0051, CRE0074, CRE0058, CRE0065, CRE0066; CRE0018, CRE0077, CRE0058, CRE0065, CRE0066; CRE0018, CRE0077, CRE0074, CRE0058, CRE0065, CRE0066; CRE0018, CRE0077, CRE0074, CRE0058, CRE0065; and CRE0051, CRE0042, wherein the CREs are present in the CRM in the recited order, and wherein they are adjacent to one another.

8 . The synthetic liver-specific CRM of claim 1 comprising a combination of CREs selected from the group consisting of:

CRE0018, CRE0051, CRE0058, CRE0065, CRE0066;

CRE0051 CRE0058, CRE0065, CRE0012;

CRE0051, CRE0058, CRE0065, CRE0066; and

CRE0051, CRE0042, wherein the CREs are present in the CRM in the recited order, and wherein they are adjacent to one another.

9 . The synthetic liver-specific CRM of claim 1 comprising a CRM selected from the group consisting of: CRM_SP0109, CRM_SP0112, CRM_SP0113, CRM SP0121, CRM SP0124, CRM SP0127, CRM SP0127A1, CRM SP0127V1, CRM_SP0127V2, CRM_SP0128, CRM_SP0131, CRM_SP0132, CRM_SP0133, CRM_SP0239, CRM_SP0240, CRM_SP0241, CRM_SP0242, CRM_SP0243, CRM_SP0244, CRM_SP0246, CRM_SP0247, CRM_SP0248, CRM_SP0249, CRM_SP0250, CRM_SP0251, CRM_SP0253, CRM_SP0254, CRM_SP0255, CRM_SP0256, CRM_SP0257, CRM_SP0258, CRM_SP0265, CRM_SP0266, CRM_SP0267, CRM_SP0268, CRM_SP0269, CRM_SP0270, CRM_SP0271, CRM_SP0272, CRM_SP0273, CRM_SP0368, CRM_SP0373, CRM_SP0378, CRM_SP0379, CRM_SP0380, CRM_SP0381, CRM_SP0384, CRM_SP0396, CRM_SP0397, CRM_SP0398, CRM_SP0403, CRM_SP0404, CRM_SP0405, CRM_SP0406, CRM_SP0407, CRM_SP0409, CRM_SP0411, CRM_SP0412, CRM_SP0413.

10 . The synthetic liver-specific CRM of claim 9 wherein the CRM is CRM_SP0239, CRM_SP0244, CRM_SP0265 or CRM_SP0412, or wherein the CRM has a length of 450, 400, 350, 300, 250, 200, 150, 100, 75, 60, 50 or fewer nucleotides.

11 . A synthetic liver-specific promoter comprising:

a) a CRM according to claim 1 operably linked to a promoter element; or

b) at least one of the following CREs;

CRE0018;

CRE0042;

CRE0051;

CRE0058;

CRE0065;

CRE0066;

CRE0068; and

CRE0074,

operably linked to a promoter element selected from CRE0059 and CRE0006.

12 . The synthetic liver-specific promoter of claim 11 comprising a combination of CREs selected from the group consisting of: CRE0051, CRE0066.2; CRE0065, CRE0051, CRE0083.1; CRE0065, CRE0066.2; CRE0066, CRE0066; CRE0065, CRE0066; CRE0074, CRE0058, CRE0065.1; CRE0051, CRE0074, CRE0058, CRE0065.1; CRE0077, CRE0074, CRE0058, CRE0065.1; CRE0078, CRE0074, CRE0058, CRE0065.1; CRE0074, CRE0074, CRE0065; CRE0058, CRE0065, CRE0066; CRE0074, CRE0058, CRE0065, CRE0066; CRE0074, CRE0058; CRE0018, CRE0051, CRE0058, CRE0065, CRE0066; CRE0051, CRE0058, CRE0065, CRE0066; CRE0051, CRE0058, CRE0065; CRE0012, CRE0051, CRE0058, CRE0065, CRE0066; CRE0051, CRE0058, CRE0065, CRE0012; CRE0051, CRE0058, CRE0065, CRE0018; CRE0051, CRE0058, CRE0065, CRE0001; CRE0051, CRE0058, CRE0065, CRE0077; CRE0051, CRE0058, CRE0018; CRE0047, CRE0051, CRE0058, CRE0065, CRE0066; CRE0051, CRE0058, CRE0065, CRE0066; CRE0077, CRE0058, CRE0065, CRE0066; CRE0078, CRE0058, CRE0065, CRE0066; CRE0051, CRE0074, CRE0058, CRE0065, CRE0066; CRE0077, CRE0074, CRE0058, CRE0065, CRE0066; CRE0078, CRE0074, CRE0058, CRE0065, CRE0066; CRE0051, CRE0074, CRE0058; CRE0077, CRE0074, CRE0058; CRE0078, CRE0074, CRE0058; CRE0051, CRE0058; CRE0068, CRE0042; CRE0051, CRE0058, CRE0066; CRE0051, CRE0065, CRE0066; CRE0018, CRE0051, CRE0074, CRE0058, CRE0065, CRE0066; CRE0018, CRE0077, CRE0058, CRE0065, CRE0066; CRE0018, CRE0077, CRE0074, CRE0058, CRE0065, CRE0066; CRE0018, CRE0077, CRE0074, CRE0058, CRE0065; and CRE0051, CRE0042,

wherein the CREs are present in the CRM in the recited order, and wherein they are adjacent to one another, or

wherein the promoter element is selected from CRE0006, CRE0059, CRE0052, CRE0079, CRE0073, or CRE0073.1.

13 . The synthetic liver-specific promoter of claim 11 comprising an individual CREs or combinations of CREs selected from the group consisting of: CRE0051, CRE0066.2; CRE0065, CRE0051, CRE0083.1; CRE0065, CRE0066.2; CRE0066, CRE0066; CRE0065, CRE0066; CRE0074, CRE0058, CRE0065.1; CRE0051, CRE0074, CRE0058, CRE0065.1; CRE0077, CRE0074, CRE0058, CRE0065.1; CRE0078, CRE0074, CRE0058, CRE0065.1; CRE0074, CRE0074, CRE0065; CRE0058, CRE0065, CRE0066; CRE0074, CRE0058, CRE0065, CRE0066; CRE0074, CRE0058; CRE0018, CRE0051, CRE0058, CRE0065, CRE0066; CRE0018; CRE0051, CRE0058, CRE0065, CRE0066; CRE0051, CRE0058, CRE0065; CRE0012, CRE0051, CRE0058, CRE0065, CRE0066; CRE0051, CRE0058, CRE0065, CRE0012; CRE0001, CRE0018; CRE0051, CRE0058, CRE0065, CRE0018; CRE0051, CRE0058, CRE0065, CRE0001; CRE0051, CRE0058, CRE0065, CRE0077; CRE0077, CRE0018; CRE0051, CRE0058, CRE0018; CRE0005, CRE0018; CRE0018, CRE0001; CRE0047, CRE0051, CRE0058, CRE0065, CRE0066; CRE0051, CRE0058, CRE0065, CRE0066; CRE0077, CRE0058, CRE0065, CRE0066; CRE0078, CRE0058, CRE0065, CRE0066; CRE0051, CRE0074, CRE0058, CRE0065, CRE0066; CRE0077, CRE0074, CRE0058, CRE0065, CRE0066; CRE0078, CRE0074, CRE0058, CRE0065, CRE0066; CRE0051, CRE0074, CRE0058; CRE0077, CRE0074, CRE0058; CRE0078, CRE0074, CRE0058; CRE0051; CRE0051, CRE0058; CRE0048, CRE0042; CRE0068, CRE0042; CRE0056, CRE0042; CRE0062, CRE0042; CRE0051, CRE0058, CRE0066; CRE0051, CRE0065, CRE0066; CRE0018, CRE0051, CRE0074, CRE0058, CRE0065, CRE0066; CRE0018, CRE0077, CRE0058, CRE0065, CRE0066; CRE0018, CRE0077, CRE0074, CRE0058, CRE0065, CRE0066; CRE0018, CRE0077, CRE0074, CRE0058, CRE0065; CRE0051, CRE0018; CRE0051, CRE0042; and CRE0042, wherein the CREs are present in the recited order, and wherein they are adjacent to one another.

14 . The synthetic liver-specific promoter according claim 11 selected from the group consisting of: SP0109, SP0112, SP0113, SP0121, SP0124, SP0127, SP0127A1, SP0127V1, SP0127V2, SP0128, SP0131, SP0132, SP0133, SP0239, SP0240, SP0241, SP0242, SP0243, SP0244, SP0246, SP0247, SP0248, SP0249, SP0250, SP0251, SP0253, SP0254, SP0255, SP0256, SP0257, SP0258, SP0265, SP0266, SP0267, SP0268, SP0269, SP0270, SP0271, SP0272, SP0273, SP0368, SP0373, SP0378, SP0379, SP0380, SP0381, SP0384, SP0396, SP0397, SP0398, SP0403, SP0404, SP0405, SP0406, SP0407, SP0409, SP0411, SP0412, and SP0413,

wherein the synthetic liver-specific promoter has length 600, 500, 450, 400, 350, 300, 250, 200, 150, 100, 75, 70 or 68 or fewer nucleotides, and/or

wherein the liver-specific promoter exhibits activity which is at least 25%, 50%, 75% or 100% of the activity of a TBG promoter, and/or

wherein the liver-specific promoter exhibits activity in non-liver cells which is 50%, 10%, 1% or less than CMV-IE.

15 . An expression cassette comprising a synthetic liver-specific promoter according to claim 11 operably linked to a sequence encoding an expression product wherein sequence encoding an expression product is a transgene encoding a therapeutic protein.

16 . A vector comprising a synthetic liver-specific CRM according to claim 1 , wherein the vector is a viral vector, an AAV vector, or a gene therapy vector.

17 . A virion comprising a vector of claim 16 .

18 . A pharmaceutical composition comprising a synthetic liver-specific CRM according to claim 1 .

19 . A isolated cell comprising a synthetic liver-specific CRM according to claim 1 , wherein the isolated cell is an isolated liver cell.

20 . A method for producing an expression product, the method comprising providing an expression cassette according to claim 15 in a liver cell and expressing an expression product from the sequence encoding the expression product.

21 . A method of expressing a therapeutic gene in a liver cell, the method comprising introducing into the liver cell an expression cassette according to claim 15 .

22 . A method of therapy of a subject in need thereof, the method comprising:

administering to the subject an expression cassette according to claim 15 .

Assignments (5)

Priority Claims (1)

Continuity (1)

References Cited (31)