Method for the construction of specific promoters
The present application relates to a system for designing promoters for selective expression of genes. Thereby identified transcription regulatory elements are selected according to a specific methodology and used to create a library of transcription regulatory elements, which are then used to construct specific promoters, especially tissue-specific promoters.
1. A vector, comprising:
a combined promoter cassette, the combined promoter cassette comprising: randomly combined elements and a minimum promoter: and
a reporter gene;
wherein each of the elements contains more than 80% sequence identity with a transcription factor regulatory element (TFRE) that is within 20 kilobases of more than fifty percent of a plurality of genes identified as being differentially expressed in a particular cell type or tissue type or under a particular condition and has a SYN value larger than 0.3, wherein the SYN value is defined as frequency (1/length) ;
wherein frequency is the frequency of occurrence of the TFRE within 20 kilobases of any of the plurality of the genes and length is the length in nucleotides of the TFRE,
such that the order of the combined elements does not occur in nature.
2. The vector of claim 1 , wherein the vector is a plasmid or viral vector.
3. The vector of claim 1 , wherein the vector is transiently expressed or integrated into a genome of a host cell.
4. The vector of claim 1 , wherein the frequency of occurrence of the TFRE within 20 kilobases of any of the plurality of the genes is the frequency of occurrence of the TFRE within 20 kilobases in the sense strand of any of the plurality of the genes.
5. The vector of claim 1 , wherein the vector does not comprise a promoter or regulatory sequences other than those present in the promoter cassette.
6. The vector of claim 1 , wherein the TFRE is associated with a plurality of genes, each of which is differentially expressed in a particular cell type or tissue type, or under a particular condition.
7. The vector of claim 6 , each of the plurality of genes is differentially expressed under a disease condition.
8. The vector of claim 7 , wherein the disease condition is cancer.
9. The vector of claim 6 , wherein each of the plurality of genes is differentially expressed under the particular condition, which is exposure to a particular biological agent, chemical agent, or microbial pathogen.
10. The vector of claim 1 , wherein the reported gene is LacZ or GFP.
11. A vector library comprising a plurality of the vectors of claim 1 .
12. The vector library of claim 11 , wherein the vector library is a retroviral vector library.
13. A cell expressing a vector of claim 1 .
14. A cell expressing a vector library of claim 11 .
15. The vector of claim 1 , wherein the vector comprises at least 3 randomly combined elements.