PLAICE DNA TRANSPOSON SYSTEM

• US 20100105140A1
• Filed: 07/16/2009
• Published: 04/29/2010
• Est. Priority Date: 07/16/2008
• Status: Abandoned Application

• Current Assignee: Recombinetics, Inc
• Original Assignee: Recombinetics, Inc
• Inventors: Leaver, Michael J., Clark, Karl J., Fahrenkrug, Scott C., Carlson, Daniel F.
• Application Number:
  US12/504,286
• Publication Number:
  US 20100105140A1
• Time in Patent Office: Days
• US Class Current: 435/455
• CPC Class Codes: C12N 9/22 Ribonucleases RNAses, DNAses catalytic nucleic acids C12N15/113

Abstract

This document describes the Passport transposon system and methods of making and using the same.

Claims

1. An isolated transposase comprising:

a polypeptide that has an amino acid sequence that has at least 80% identity to SEQ ID NO;

29, with the polypeptide specifically binding to the a nucleic acid fragment that comprises an inverted terminal repeat sequence of at least one of SEQ ID NO;

34 and SEQ ID NO;

35, with the polypeptide be capable of catalyzing the integration of a target nucleic acid into a vertebrate cell.

2. The transposase of claim 1 wherein the target nucleic acid is integrated into the genome of the vertebrate cell.

3. The transposase of claim 1 comprising SEQ ID NO:

29.

4. A nucleic acid encoding the transposase of claim 1.

5. The nucleic acid of claim 4, wherein the nucleic acid is a portion of a plasmid.

6. A cell comprising the transposase of claim 1.

7. A transposon comprising a first nucleic acid fragment with at least 80% identity to SEQ ID NO:

34 and a second nucleic acid fragment with at least 80% identity to SEQ ID NO;

35, wherein the transposon can be specifically bound by a polypeptide having the sequence of SEQ ID NO;

29.

8. The transposon of claim 7 further comprising a target nucleic acid fragment that is located between the first nucleic acid fragment and the second nucleic acid fragment, wherein the target nucleic acid is mobilizable by the polypeptide to be integrated into the genome of a vertebrate cell.

9. A cell comprising the transposon of claim 8.

10. A gene transfer system to introduce DNA into the DNA of a cell comprising:

a transposase or a nucleic acid encoding a transposase, with the transposase having a sequence with at least 80% identity to SEQ ID NO;

29; and

a transposon that comprises a target nucleic acid that is mobilizable by the transposase into a genome of a vertebrate cell.

11. The system of claim 10 wherein the transposon comprises SEQ ID NO:

34 and SEQ ID NO;

35.

12. The system of claim 10 wherein the target nucleic acid further comprises a promoter.

13. The system of claim 10 wherein the transposase has the sequence of SEQ ID NO:

29.

14. The system of claim 10 wherein the transposase is provided as RNA.

15. The system of claim 10 wherein the transposase is provided as a polypeptide.

16. The system of claim 10 wherein the transposase is provided as the nucleic acid encoding a transposase and is part of a plasmid.

17. A cell comprising:

a transposase or a nucleic acid encoding a transposase, with the transposase having a sequence with at least 80% identity to SEQ ID NO;

29; and

a transposon that comprises a target nucleic acid that is mobilizable by the transposase into a genome of a vertebrate cell.

18. The cell of claim 17 further comprising the target nucleic acid mobilized into the genome of the cell.

19. A method of introducing a target nucleic acid into DNA in a cell comprising:

introducing into the cell

(a) a polypeptide that has an amino acid sequence that has at least 80% identity to SEQ ID NO;

29, with the polypeptide possessing binding to a nucleic acid fragment that comprises an inverted terminal repeat sequence of at least one of SEQ ID NO;

34 and SEQ ID NO;

35, with the polypeptide catalyzing the integration of a target nucleic acid into a vertebrate cell, or

(b) a transposase or a nucleic acid encoding a transposase, with the transposase having a sequence with at least 80% identity to SEQ ID NO;

29; and

a transposon that comprises a target nucleic acid that is mobilizable by the transposase into a genome of a vertebrate cell, or

the polypeptide of (a) and the transposase or nucleic acid of (b).