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US 8,137,974 B2Granted
Transposition of maize AC/DS elements in vertebrates
AssigneeTemasek Life Sciences Laboratory Limited
InventorsSergey Parinov, Alexander Emelyanov
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Abstract
The present invention is directed to the use of the maize Ac/Ds transposable elements in vertebrates.
Claims31
A method of integrating a polynucleotide into the DNA of a vertebrate cell comprising:
- introducing into the vertebrate cell a modified Ac transposase or a nucleic acid encoding the modified Ac transposase, wherein the modified Ac transposase comprises (i) a nuclear localization signal (NLS) and (ii) a truncated Ac transposase, wherein the NLS is fused to the N-terminus of the truncated Ac transposase, wherein the NLS localizes a transposition reaction to the nucleus of the vertebrate cell and wherein the truncated Ac transposase comprises the amino acid sequence set forth in SEQ ID NO:8 and catalyzes the integration of the polynucleotide into the DNA of the vertebrate cell, wherein the NLS comprises the amino acid sequence set forth in SEQ ID NO:2 or the amino acid sequence set forth in SEQ ID NO:9;
- introducing into the vertebrate cell a nucleic acid fragment comprising the polynucleotide positioned between 5′ end and 3′ end cis-required sequences of a Ds element that bind to the modified Ac transposase;
- and
- integrating the polynucleotide into the DNA of the vertebrate cell.
1 more element
Dependencies are inferred from the claim text.