Liver-specific constructs and methods of use thereof

Patent metadata

Field	Value
Publication number	US-10828376-B2
Application number	US-201816164346-A
Country	US
Kind code	B2
Filing date	Oct 18, 2018
Priority date	Oct 28, 2015
Publication date	Nov 10, 2020
Grant date	Nov 10, 2020

Abstract

Described herein are constructs used for liver-specific expression of a transgene.

First claim

What is claimed is:

1. A polynucleotide expression construct comprising at least one spacer sequence comprising an insulator sequence, a liver-specific enhancer sequence, a promoter sequence and a transgene, wherein the insulator sequence is selected from the group consisting of SEQ ID NOs:28, 29, 30 and 38.

2. The polynucleotide expression construct of claim 1 , further comprising an intron sequence, wherein the enhancer sequence comprises a wild-type or mutated Serpin 1 enhancer; and/or the promoter sequence comprises a wild-type or mutated transthyretin (TTR) promoter; and/or the intron sequence comprises a wild-type or mutated minute virus of mice (MVM) intron sequence.

3. The polynucleotide expression construct of claim 2 , comprising at least two spacer sequences flanking the liver-specific enhancer sequence, the promoter sequence, the intron sequence and the transgene.

4. The polynucleotide expression construct of claim 1 , wherein the transgene encodes a protein lacking or deficient in a hemophilia or a lysosomal storage disease.

5. The polynucleotide expression construct of claim 4 , wherein the transgene further encodes one or more nucleases.

6. An AAV vector comprising the polynucleotide expression construct of claim 1 , wherein the polynucleotide expression construct is between 5′ and 3′ inverted terminal repeats (ITRs) of the AAV vector.

7. A pharmaceutical composition comprising the AAV vector according to claim 6 and a pharmaceutically acceptable carrier.

8. A kit comprising the polynucleotide expression construct according to claim 1 .

9. The polynucleotide expression construct of claim 1 , comprising in the following order: the sequence of SEQ ID NO:28, the liver-specific enhancer sequence, the promoter sequence, the transgene, and the sequence of SEQ ID NO:30.

10. The polynucleotide expression construct of claim 2 , comprising in the following order: the sequence of SEQ ID NO:28, the liver-specific enhancer sequence, the promoter sequence, the intron sequence, the transgene, a polyadenylation sequence and the sequence of SEQ ID NO:30.

11. The polynucleotide expression construct of claim 1 , comprising at least one spacer sequence comprising an insulator sequence, wherein the insulator sequence is SEQ ID NO:28.

12. The polynucleotide expression construct of claim 1 , comprising at least one spacer sequence comprising an insulator sequence, wherein the insulator sequence is SEQ ID NO:30.

13. The polynucleotide expression construct of claim 1 , comprising a first spacer sequence comprising SEQ ID NO:28 and a second spacer sequence comprising SEQ ID NO:30.

Assignees

• Sangamo Therapeutics Inc

Inventors

• Riley Brigit E
• Lee Gary K

Classifications

• C12N2830/60

  from viruses · CPC title

• C12N2830/42

  being an intron or intervening sequence for splicing and/or stability of RNA · CPC title

• C12N2830/008

  cell type or tissue specific enhancer/promoter combination · CPC title

• C12N2750/14145

  Special targeting system for viral vectors · CPC title

• C12N2750/14143

  viral genome or elements thereof as genetic vector · CPC title