Expression systems
View Patent ↗A gene expression system is provided. The system comprises at least one coding sequence to be expressed in an organism, and at least one promoter operably linked thereto. It further comprises at least one splice control sequence which, in cooperation with a spliceosome, mediates alternative splicing of RNA transcripts of the coding sequence. The mediation of alternative splicing is in a sex-specific, stage-specific, germline-specific and tissue-specific manner.
1. A gene expression system comprising a coding sequence to be expressed in an insect, a promoter operably linked thereto, and an intronic splice control sequence, which is flanked on its 5′ end by a guanine (G) nucleotide,
wherein said intronic splice control sequence, in cooperation with a spliceosome, is capable of sex-specifically mediating alternative splicing of RNA transcripts of the coding sequence, the coding sequence encodes a protein having a lethal, deleterious or sterilizing effect and is not a sequence associated with or linked to the intronic splice control sequence in a native context,
wherein the intronic splice control sequence is obtained from AaActin-4, Dsx, Bztra or Cctra, and
wherein the insect is selected from the group consisting of mosquito, bollworm and medfly.
2. The gene expression system of claim 1 , further comprising a coding sequence encoding a marker.
3. The gene expression system of claim 1 , wherein a product of the coding sequence serves as a positive transcriptional control factor for the promoter, whereby the product, or the expression of the product, is controllable.
4. The gene expression system of claim 1 , wherein the protein is an apoptosis-inducing factor.
5. The gene expression system of claim 4 , wherein the protein is selected from the group consisting of: AIF, Hid, and Reaper (Rpr).
6. The gene expression system of claim 1 , wherein the protein has a lethal effect.
7. The gene expression system of claim 1 , wherein the intronic splice control sequence is obtained from Dsx and the expression system comprises a sequence selected from the group consisting of: SEQ ID NO. 40, SEQ ID NO. 41, SEQ ID NO. 42, SEQ ID NO. 43, SEQ ID NO. 44, SEQ ID NO. 45, and SEQ ID NO. 71.
8. The gene expression system of claim 1 , wherein the intronic splice control sequence is obtained from Dsx, the expression system comprising a construct selected from the group consisting of: pLA3435(SEQ ID NO. 46), pLA3359 (SEQ ID NO. 47), pLA3433 (SEQ ID NO. 48).
9. The gene expression system of claim 1 , wherein the intronic splice control sequence is obtained from Cctra, the expression system comprising a construct selected from the group consisting of:: pLA1188 (SEQ ID NO. 49), pLA3077 (SEQ ID NO. 50), pLA3097(SEQ ID NO. 51), pLA3233 (SEQ ID NO. 52), pLA3014 (SEQ ID NO 53), pLA3166 (SEQ ID NO. 54), and pLA3242 (SEQ ID NO. 56).
10. The gene expression system of claim 1 , wherein the intronic splice control sequence is obtained from Bztra, the expression system comprising a construct set forth in pLA3376 (SEQ ID NO. 55).
11. The gene expression system of claim 1 , wherein the protein is a ubiquitin protein which has the sequence according to SEQ ID NO. 73.
12. The gene expression system of claim 1 , comprising two or more splice control sequences.