Self-inactivating transposase plasmids and uses thereof
Some embodiments provided herein relate to gene delivery systems and methods using a single plasmid that carries a self-inactivating transposase gene and a corresponding transposon. Some embodiments include nucleic acids having certain sequences, vector including such nucleic acids, and compositions including the vectors.
1 . A vector for delivery of a payload into a cell genome, the vector comprising (a), (b), and (c) in a 5′ to 3′ order:
(a) a first nucleic acid comprising (i) a nucleotide sequence encoding a transposase, and (ii) an intron comprising a first transposase recognition site (TRS), wherein the intron is inserted into the nucleotide sequence encoding the transposase;
(b) a second nucleic acid encoding the payload; and
(c) a second TRS.
2 . The vector of claim 1 , wherein the vector is a circular vector.
3 . The vector of claim 1 , wherein the transposase is a Tn5 transposase, a MuA transposase, a Tn552 transposase, or a mariner transposase.
4 . The vector of claim 1 , wherein the transposase is encoded by a sequence having at least 90% sequence identity to the nucleotide sequence of SEQ ID NO:01 or SEQ ID NO:03.
5 . The vector of claim 1 , wherein the intron is derived from a nucleic acid lacking the first TRS and, wherein the nucleic acid lacking the first TRS comprises a sequence having at least 90% sequence identity to the nucleotide sequence of SEQ ID NO:02.
6 . The vector of claim 1 , wherein the first TRS and the second TRS are both derived from a transposon selected from a Tn5 transposon, a MuA transposon, a Tn552 transposon, or a mariner transposon.
7 . The vector of claim 1 , wherein the first TRS and the second TRS comprise a sequence having at least 90% sequence identity to the nucleotide sequence of SEQ ID NO:05 or SEQ ID NO:13.
8 . The vector of claim 1 , wherein the payload encodes a therapeutic protein.
9 . The vector of claim 1 , further comprising a first promoter operably linked to the first nucleic acid.
10 . The vector of claim 1 , further comprising a selectable marker.
11 . The vector of claim 10 , wherein the selectable marker encodes a dihydrofolate reductase (DHFR).
12 . The vector of claim 1 , further comprising a nucleic acid encoding a first polyA signal operably linked to the second nucleic acid, or a second polyA signal operably linked to the third nucleic acid.
13 . A cell comprising the vector of claim 1 .
14 . The cell of claim 13 , wherein the cell is a mammalian cell.
15 . The cell of claim 14 , wherein the cell is a human cell.