Nucleic acid construct and use of the same
The present invention is related to a nucleic acid construct comprising an expression unit for the expression of E1B, wherein the expression unit comprises a promoter, a nucleotide sequence coding for E1B, and a 3′UTR, wherein the promoter is operatively linked to the nucleotide sequence coding for E1B, wherein the 3′UTR comprises 30 or less than 30 Exonic Enhancer Elements (ESEs), preferably 20 or less than 20 Exonic Enhancer Elements (ESEs), and wherein the 3′ UTR is a non-viral 3′ UTR.
1. A nucleic acid construct comprising
(i) an expression unit for the expression of E1 B, wherein the expression unit comprises a promoter, a nucleotide sequence coding for E1 B, and a 3′ UTR, wherein the promoter is operatively linked to the nucleotide sequence coding for E1 B, wherein the 3′ UTR comprises 30 or fewer than 30 Exonic Splicing Enhancer elements (ESEs), and wherein the 3′ UTR is a non-viral 3′ UTR, and
(ii) an expression unit for the expression of E1A, wherein the expression unit comprises a promoter, a nucleotide sequence coding for E1A, and a 3′ UTR,
wherein the promoter is operatively linked to the nucleotide sequence coding for E1A, wherein the expression unit for the expression of E1A and the expression unit for the expression of EIB form a combined expression unit, wherein the combined expression unit comprises the nucleotide sequence of SEQ ID NO: 9 or SEQ ID NO: 15.
2. The nucleic acid construct of claim 1 , wherein the 3′ UTR comprises 20 or fewer than 20 Exonic Splicing Enhancer elements (ESEs).
3. The nucleic acid construct of claim 1 , wherein the nucleic acid construct is a single nucleic acid molecule comprising both the expression unit for the expression of E1A and the expression unit for the expression of E1 B.
4. The nucleic acid construct according to claim 1 , wherein the Exonic Splicing Enhancer elements (ESEs) are contained within a stretch of nucleotides of the 3′ UTR of the expression unit for the expression of E1 B, and wherein said stretch of nucleotides comprises the 200 nucleotides of the 5′ end of the 3′ UTR of the expression unit for the expression of E1 B.
5. The nucleic acid construct according to claim 1 , wherein the 3′ UTR of the expression unit for the expression of E1 B is a mammalian 3′ UTR.
6. The nucleic acid construct according to claim 1 , wherein the expression unit for the expression of E1A and the expression unit for the expression of E1 B are arranged in a 5′->3′ direction in the nucleic acid construct as follows: the promoter of the expression unit for the expression of E1A, the nucleotide sequence coding for E1A and the 3′ UTR, the promoter of the expression unit for the expression of E1 B, the nucleotide sequence coding for E1 B, a splice donor site, an intron, a splice acceptor site and the 3′ UTR.
7. The nucleic acid construct according to claim 1 , wherein the nucleic acid construct is coding for and capable of expressing E1A, E1 B 55K, E1 B 19K and/or E1 B 84R.
8. The nucleic acid construct according to claim 7 , wherein the nucleic acid construct is capable of expressing E1A, E1 B 55K, and E1 B 19K.
9. A vector comprising the nucleic acid construct according to claim 1 .
10. A cell comprising a the vector according to claim 9 .
11. The cell according to claim 10 , wherein the cell is an amniocytic cell line.
12. The nucleic acid construct according to claim 7 , wherein the nucleic acid construct is capable of expressing E1A, E1 B 55 K, E1 B 19 K and E1 B 84R.