Transposon for genome manipulation
OwnerChunxiao WUinventor
Life of this patent
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Abstract
Disclosed is a vector comprising a piggyBac like element (PLE) transposon and/or transposase. Also disclosed are vectors comprising a combination of the vectors of the present disclosure, a transformation system comprising the vectors of the present disclosure, an oligonucleotide primer, methods of amplifying nucleotide sequences, methods of transforming a cell with a gene and a transgenic organism.
Claims41
A vector, comprising a piggyBac like element (PLE) transposon.
(canceled)
A vector, comprising a nucleotide sequence encoding a piggyBac like element (PLE) transposase.
(canceled)
20-27. (canceled)
A transformation system, comprising
- a) a first vector, comprising a piggyBac like element (PLE) transposon;
- and
- a second vector, comprising a nucleotide sequence encoding a piggyBac like element (PLE) transposase;
- or
- b) a single vector comprising the piggyBac like element transposon and the nucleotide sequence that encodes the piggyBac like element transposase.
31-40. (canceled)
A kit for transforming a cell, comprising one or more vectors, wherein each vector comprises piggyBac like element (PLE) transposon.
44-46. (canceled)
A kit for transforming a cell, comprising a single vector comprising a piggyBac like element transposon and a nucleotide sequence that encodes the piggyBac like element transposase.
(canceled)
A method of amplifying a nucleotide sequence of any one of SEQ ID NOs:
- 1, 3, 4 or 5, wherein the amplification of SEQ ID NO:
- 1 comprises performing a polymerase chain reaction using a forward primer comprising the nucleotide sequence of SEQ ID NO:
- 6 and a reverse primer comprising the nucleotide sequence of SEQ ID NO:
- 7;
- wherein the amplification of SEQ ID NO:
- 3 comprises performing a polymerase chain reaction using a forward primer comprising nucleotide sequence of SEQ ID NO:
- 8 and a reverse primer comprising nucleotide sequence of SEQ ID NO:
- 9;
- wherein the amplification of SEQ ID NO:
- 4 comprises performing a polymerase chain reaction using a forward primer comprising nucleotide sequence of SEQ ID NO:
- 10 and a reverse primer comprising nucleotide sequence of SEQ ID NO:
- 11 or SEQ ID NO:
- 20;
- wherein the amplification of SEQ ID NO:
- 5 comprises performing a polymerase chain reaction using a forward primer comprising nucleotide sequence of SEQ ID NO:
- 12 and a reverse primer comprising nucleotide sequence of SEQ ID NO:
- 13.
50-52. (canceled)
A method of transforming a cell with a gene, comprising:
- a) transfecting the cell with a first vector, comprising a piggyBac like element (PLE) transposon, wherein the first vector comprises the 5′ and 3′ ends of a piggyBac like element (PLE) transposon;
- and
- transfecting the cell with a second vector, comprising a nucleotide sequence encoding a piggyBac like element (PLE) transposase, wherein the second vector comprises a nucleotide sequence that encodes a piggyBac like element transposase;
- or
- b) transfecting the cell with a single vector, wherein the single vector comprises the 5′ and 3′ ends of the piggyBac like element transposon and the nucleotide sequence that encodes the PLE transposase.
(canceled)
(canceled)
A method of transforming a cell with a gene, comprising:
- transfecting the cell with a first vector, wherein the first vector comprises the 5′ and 3′ ends of a piggyBac like element (PLE) transposon;
- and
- delivering a nucleic acid encoding a nucleotide sequence encoding a piggyBac like element (PLE) transposase or a transposase protein either together with the first vector or before transfection of the first vector or after transfection of the first vector.
A method of transforming a cell with a gene, comprising:
- delivering a nucleic acid encoding a nucleotide sequence encoding a piggyBac like element (PLE) transposase or a transposase protein into the cell, wherein the cell already comprises the 5′ and 3′ ends of a piggyBac like element (PLE) transposon.
59-61. (canceled)
A non-human transgenic organism comprising cells which have been transformed by a method of transforming a cell with a gene, comprising:
- a) transfecting the cell with a first vector, comprising a piggyBac like element (PLE) transposon, wherein the first vector comprises the 5′ and 3′ ends of a piggyBac like element (PLE) transposon;
- and transfecting the cell with a second vector, comprising a nucleotide sequence encoding a piggyBac like element (PLE) transposase, wherein the second vector comprises a nucleotide sequence that encodes a piggyBac like element transposase;
- or
- b) transfecting the cell with a single vector, wherein the single vector comprises the 5′ and 3′ ends of the of piggyBac like element transposon and the nucleotide sequence that encodes the PLE transposase.
63-65. (canceled)
Dependencies are inferred from the claim text.