Methods and compositions for regulation of transgene expression

Patent metadata

Field	Value
Publication number	US-10975375-B2
Application number	US-201715690067-A
Country	US
Kind code	B2
Filing date	Aug 29, 2017
Priority date	Sep 21, 2011
Publication date	Apr 13, 2021
Grant date	Apr 13, 2021

Abstract

Nucleases and methods of using these nucleases for expressing a transgene from a safe harbor locus in a secretory tissue, and clones and animals derived therefrom.

First claim

What is claimed is:

1. A method of integrating a transgene into a human or mouse liver cell, the method comprising:
administering a polynucleotide encoding a nuclease to the liver cell, the nuclease comprising (i) a DNA-binding domain that binds to a target site in an endogenous albumin gene, wherein the DNA-binding domain is a zinc finger protein (ZFP) comprising four, five or six zinc finger domains ordered F1 to F4, F1 to F5, or F1 to F6, each zinc finger domain comprising a recognition helix region, wherein the recognition helix regions are ordered as shown in a single row of the following Table:

F1
F2
F3
F4
F5
F6

TSGSLTR
RSDALST
QSATRTK
TSGHLSR
QSGNLAR
NA

(SEQ ID
(SEQ ID
(SEQ ID
(SEQ ID
(SEQ ID

NO:1)
NO:2)
NO:3)
NO:4)
NO:5)

RSDHLSA
TKSNRTK
DRSNLSR
WRSSLRA
DSSDRKK
NA

(SEQ ID
(SEQ ID
(SEQ ID
(SEQ ID
(SEQ ID

NO:6)
NO:7)
NO:8)
NO:9)
NO:10)

TSGNLTR
DRSTRRQ
TSGSLTR
ERGTLAR
TSANLSR
NA

(SEQ ID
(SEQ ID
(SEQ ID
(SEQ ID
(SEQ ID

NO:11)
NO:12)
NO:1)
NO:13)
NO:14)

DRSALAR
RSDHLSE
HRSDRTR
QSGALAR
QSGHLSR
NS

(SEQ ID
(SEQ ID
(SEQ ID
(SEQ ID
(SEQ ID

NO:15)
NO:16)
NO:17)
NO:18)
NO:19)

RSDNLST
DRSALAR
DRSNLSR
DGRNLRH
RSDNLAR
QSNALNR

(SEQ ID
(SEQ ID
(SEQ ID
(SEQ ID
(SEQ ID
(SEQ ID

NO:20)
NO:15)
NO:8)
NO:21)
NO:22)
NO:23)

DRSNLSR
LKQVLVR
QSGNLAR
QSTPLFA
QSGALAR
NA

(SEQ ID
(SEQ ID
(SEQ ID
(SEQ ID
(SEQ ID

NO:8)
NO:24)
NO:5)
NO:25)
NO:18)

DRSNLSR
DGRNLRH
RSDNLAR
QSNALNR
NA
NA

(SEQ ID
(SEQ ID
(SEQ ID
(SEQ ID

NO:8)
NO:21)
NO:22)
NO:23)

RSDNLSV
HSNARKT
RSDSLSA
QSGNLAR
RSDSLSV
QSGHLSR

(SEQ ID
(SEQ ID
(SEQ ID
(SEQ ID
(SEQ ID
(SEQ ID

NO:26)
NO:27)
NO:28)
NO:5)
NO:29)
NO:19)

RSDNLSE
ERANRNS
QSANRTK
ERGTLAR
RSDALTQ
NA

(SEQ ID
(SEQ ID
(SEQ ID
(SEQ ID
(SEQ ID

NO:30)
NO:31)
NO:32)
NO:13)
NO:33)

TSGSLTR
DRSNLSR
DGRNLRH
ERGTLAR
RSDALTQ
NA

(SEQ ID
(SEQ ID
(SEQ ID
(SEQ ID
(SEQ ID

NO:1)
NO:8)
NO:21)
NO:13)
NO:33)

QSGHLAR
RSDHLTQ
RSDHLSQ
WRSSLVA
RSDVLSE
RNQHRKT

(SEQ ID
(SEQ ID
(SEQ ID
(SEQ ID
(SEQ ID
(SEQ ID

NO:34)
NO:35)
NO:36)
NO:37)
NO:38)
NO:39)

QSGDLTR
RSDALAR
QSGDLTR
RRDPLIN
RSDNLSV
IRSTLRD

(SEQ ID
(SEQ ID
(SEQ ID
(SEQ ID
(SEQ ID
(SEQ ID

NO:40)
NO:41)
NO:40)
NO:42)
NO:26)
NO:43)

RSDNLSV

Assignees

• Sangamo Therapeutics Inc

Inventors

• Rebar Edward J

Classifications

• C12N15/62 Primary

  DNA sequences coding for fusion proteins · CPC title

• C12N2710/10041

  Use of virus, viral particle or viral elements as a vector · CPC title

• C12N9/96

  Stabilising an enzyme by forming an adduct or a composition; Forming enzyme conjugates · CPC title

• C12N9/22 Primary

  Ribonucleases {[RNase]; Deoxyribonucleases [DNase]} · CPC title

• C12N5/067

  Hepatocytes · CPC title