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Revision as of 11:12, 4 October 2022

CMV promoter

Part Description

CMV (cytomegalovirus) promoter is a strong constitutive promoter commonly used in mammalian expression systems to produce high levels of expression, However CMV promoter is on of the most variable types of promoter as it is highly variable between different cell types such as being strong in some cell types and being much weaker in other types

Usage

We used a mammalian expression systems to regulate the expression of Cas12g which have a remarkable role on tuning and controlling our circuit activity and the levels of PAH to be constant within normal level to avoid overexpression of cas12g

Literature Characterization

An essential part originated from the human cytomegalovirus we employ it to regulate the expression of the Cas12g the protein domain of our CRISPR regulatory system CMV promoter is known to have a moderate level of expression lesser than other promoters we take advantage of that to limit the possibility of overexpression of Cas12g to reduce the off targeting effect

Figure.1 Effect of different promoter on transfection efficiency and transient transgene expression using eGFP antibody analysis.

The greatest expression levels were seen in the cells transfected with CHEF-1 promoter-carrying vectors, followed by those with HEF-1, CMV mutant, CMV, mouse CMV, CAG, CAG enhancer, and PGK. When the amount of eGFP expression under the CMV promoter was taken into account as 100, the levels of eGFP expression under the PGK, CHEF-1, HEF-1, mouse CMV, CMV mutant, CAG, and CAG enhancer promoters were 218.13, 184.21, 164.33, 49.12, 47.95, and 8.77, respectively.

References

1. Pham, P. L., Kamen, A. & Durocher, Y. Large-scale transfection of mammalian cells for the fast production of recombinant protein. Mol. Biotechnol. 34, 225–237 (2006).

Sequence and Features