A CCT Promoter to Enhance Protein Expression
- Detailed Technology Description
- Inventor:Rama K. MallampalliProfessor of MedicineChief, Pulmonary Division of VA Pittsburgh Health System
- *Abstract
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Background
A promoter serves to regulate the expression of an endogenous or transfected gene. More specifically, the promoter serves as the first level of regulation controlling the amount of protein that is made by the gene. In research and clinical applications, desirable traits for a promoter include a compact size, which leaves more room for coding sequence and can increase transfection efficiency, as well as a strong constitutive or inducible level of expression, which produces more of the target protein in desired locations.
Technology
Researchers at the University of Iowa have isolated approximately 200 base-pairs upstream of the CCT gene transcriptional start site. This short sequence serves as a strong promoter for research and clinical gene expression applications. Several transcriptional binding sites have been identified in this sequence including sites for CREB, NFkappaB, and SRE. SRE has been shown to give the promoter the additional characteristic of an inducible increase in expression of the target gene. The basal level of transcription has been shown to be very strong in several cell types and can be significantly augmented through the additional of all-trans retinoic acid. Additionally, a mutated sequence has been created that eliminates the SRE and the inducibility associated with it.
Advantages
COMPACT SEQUENCE. The entire promoter sequence consists of slightly over 200 base pairs in length.
STRONG BASAL EXPRESSION. Basal expression is consistently greater than that of SV40 promoter and has been shown to be greater than CMV promoter, as well.
INCLUDES INDUCIBLE ELEMENTS. A sterol response element (SRE) has been identified at the 5' end of the sequence, which can be utilized for induction of expression. In addition, All-trans retinoic acid treatment has been shown in studies to significantly upregulate the expression via the CCT promoter. A mutated sequence that no longer contains the SRE has also been created. Applications of this promoter include the generation of recombinant proteins, reporter gene expression, and incorporation of vectors used in gene delivery and immunization protocols.
Patent Information US 7,230,091
- *Licensing
- Catherine S. KohSenior Licensing AssociateUniversity of Iowa Research FoundationPh: (319) 335-4659 catherine-koh@uiowa.edu
- Country/Region
- USA
