IP Library › Granted Patent US 7,312,075
Granted Patent B1
US 7,312,075 · App. 10/030,658 · Granted Dec 25, 2007

Trap vectors and gene trapping by using the same

Assignee: Transgenic Inc.
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Quick Facts
Patent No.
US 7,312,075
App. No.
10/030,658
Granted
Dec 25, 2007
Kind
B1
Abstract

A trap vector containing a loxP sequence composed of inverted repeat sequence 1, a spacer sequence and inverted repeat sequence 2 in this order, the loxP sequence being a mutant loxP wherein a part of the inverted repeat sequence 1 or 2 is mutated.

Claims (29)

1. A trap vector selected from the group consisting of the following (a) to (i):

(a) SP-SA-lox71-IRES-M-loxP-PV-SP;

(b) SP-lox71-IRES-M-loxP-PV-SP;

(c) SP-lox71-IRES-M-loxP-pA-PV-SP;

(d) SP-lox71-IRES-M-loxP-puro-pA-PV-SP;

(e) lox71-M-loxP-pA-lox2272-PV-lox511;

(f) lox71-IRES-M-loxP-pA-lox2272-PV-lox511;

(g) (lox71-integrated SA)-M-loxP-pA-lox2272-PV-lox511;

(h) (lox71-integrated SA)-IRES-M-loxP-pA-loxP-2272-PV-lox511; and

(i) (lox71-integrated SA)-M-loxP-pA-lox2272-promote-M-lox511-SD;

wherein SP represents any sequence; SA represents a splice acceptor; SD represents a splice don or; IRES represents an internal ribosomal entry site; M represents a marker gene; puro represents puromycin resistance gene; pA represents a poly(A) sequence, and PV represents a plasmid vector.

2. The trap vector of claim 1 , wherein the plasmid vector is any one selected from the group consisting of pBR, pUC, pSP and pGEM.

3. A trap vector selected from the group consisting of the following (a) to (i):

(a) SP-SA-lox66-IRES-M-loxP-PV-SP;

(b) SP-lox66-IRES-M-loxP-PV-SP;

(c) SP-lox66-IRES-M-loxP-pA-PV-SP;

(d) SP-lox66-IRES-M-loxP-puro-pA-PV-SP;

(e) lox66-M-loxP-pA-lox2272-PV-lox511;

(f) lox66-IRES-M-loxP-pA-lox2272-PV-lox511;

(g) (lox66-integrated SA)-M-loxP-pA-lox2272-PV-lox511;

(h) (lox66-integrated SA)-IRES-M-loxP-pA-loxP-2272-PV-lox511; and

(i) (lox66-integrated SA)-M-loxP-pA-lox2272-promote-M-lox511-SD;

wherein SP represents any sequence; SA represents a splice acceptor; SD represents a splice donor; IRES represents an internal ribosomal entry site; M represents a marker gene; puro represents puromycin resistance gene; pA represents a poly(A) sequence; and PV represents a plasmid vector.

4. A method of gene trapping, comprising the steps of:

introducing the trap vector of any one of claims 1 , 2 or 3 into embryonic stem cells;

culturing the embryonic stem cells;

selecting those cells which exhibit a pattern of single copy integration of the trap vector; and

isolating the trapped gene.

5. Embryonic stem cells comprising the trap vector of claims 1 , 2 , or 3 .

Assignments (2)
CHANGE OF ADDRESS OF ASSIGNEE Recorded Mar 10, 2017
From: TRANS GENIC INC.
To: TRANS GENIC INC.
Reel/Frame 041962/0995 →
ASSIGNMENT OF ASSIGNOR'S INTEREST Recorded Jan 11, 2002
From: YAMAMURA, KEN-ICHI; ARAKI, KIMI
To: TRANSGENIC INC.
Reel/Frame 012827/0809 →
Priority Claims (1)
JP 11-200997 · Jul 14, 1999 · national