In vivo genome-wide mutagenesis

• US 8,546,135 B2
• Filed: 02/08/2008
• Issued: 10/01/2013
• Est. Priority Date: 02/09/2007
• Status: Expired due to Fees

• Current Assignee: University of Utah Research Foundation (State of Utah)
• Original Assignee: University of Utah Research Foundation (State of Utah)
• Inventors: Wu, Sen, Capecchi, Mario R.
• Primary Examiner(s): DUNSTON, JENNIFER ANN
• Application Number:
  US12/522,075
• Publication Number:
  US 20100178696A1
• Time in Patent Office: 2,062 Days
• Field of Search: None
• US Class Current: 435/320.1
• CPC Class Codes: C12N 15/1051 Gene trapping, e.g. exon-, intron-, IRES-, signal sequence-trap cloning, trap vectors
• CPC Class Codes: C12N 15/8509 for producing genetically modified animals, e.g. transgenic
• CPC Class Codes: C12N 2800/30 Vector systems comprising sequences for excision in presence of a recombinase, e.g. loxP or FRT
• CPC Class Codes: C12N 2800/90 Vectors containing a transposable element
• CPC Class Codes: C12N 2840/203 having an IRES

Abstract

Disclosed herein are compositions and methods for deleting or duplicating DNA in a mammalian genome. Also disclosed are compositions and methods for generating a random genome-wide chromosome rearrangement. Also disclosed are compositions and methods for streamlined construction of gene targeting vectors.

Claims

1. A gene-trap vector, comprising the formula:

3′

TR-lox-SA-R¹—

X—

R²-lox-X-5′

TR

wherein 3′

TR and 5′

TR are piggyBac 3′ and

5′

transposon terminal sequences comprising 3′ and

5′

inverted terminal repeats (ITR), respectively;

wherein the lox sites are in the same orientation;

wherein SA is a splice acceptor;

wherein R¹ is a first reporter sequence linked to SA; and

wherein R² is a second reporter sequence functionally linked to an expression control sequence, and wherein X is a recombination site, wherein X is FRT, attB or attP.

2. A gene-trap vector, comprising the formula:

3′

TR-lox-SA-R¹—

X—

R²—

X-lox-5′

TR

wherein 3′

TR and 5′

TR are piggyBac 3′ and

5′

transposon terminal sequences comprising 3′ and

5′

inverted terminal repeats (ITR), respectively;

wherein the lox sites are in the same orientation;

wherein SA is a splice acceptor;

wherein R¹ is a first reporter sequence linked to SA; and

wherein R² is a second reporter sequence functionally linked to an expression control sequence, and wherein X is a recombination site, wherein X is FRT, attB or attP.

3. A gene-trap vector comprising the nucleic acid sequence set forth in SEQ ID NO:

105, SEQ ID NO;

106, or SEQ ID NO;

107.