Difference between revisions of "Part:BBa K4586016"

Revision as of 15:45, 5 October 2023

CMV

Part Description

CMV promoter is derived from human Cytomegalovirus, which belongs to Herpesvirus group. All family members share the ability to remain in latent stage in the human body. CMV is located upstream of immediate-early gene. However, CMV promoter is an example of widely used promoters and is present in mammalian expression vectors. The advantage of CMV is the high-level constitutive expression in mostly all human tissues. Due to its high transcription levels, it is a potent promoter that is employed to induce gene expression.

Usage

It is a promoter that controls the expression of CD63-L7Ae, the syn-notch receptor, and booster genes.

Literature Characterization

The study used Genomatix software for analysis of the CMV promoter (-550 to +48 relative to the transcription start site; TSS) for the presence of putative transcription factor regulatory elements (TFREs). They recognized one hundred and eight discrete TFREs in the CMV promoter for the presence of their cognate transcription factors (TFs) in HEK293 cells.

(A) THEY MADE RNA-seq analysis of the HEK293 cell transcriptome to assess the gene expression level of TFs. Points represent the expression level of each TF sampled at exponential and stationary phases of culture. They thought that genes with more than two transcripts per million (log2 TPM > 1) are actively transcribed genes. (B) They chose a CMV promoter sequence with 25 selected TFREs for in vitro analysis. The TSS is referred to with an arrow.

charactrizion by mathematical modeling

That part plays an important role for expression of the synthetic notch receptors that bind to BCR so we used a set of ordinary differential equations (ODEs) and plotted them in order to build a model that describes the binding between the external domain (CCP1) of synthetic notch receptor and BCR.

Graph(1) shows an increase in the free portion of CCP1 of synthetic notch (represented as blue line) then decreases as BCR binds to it. As the binding occurs, BCR decreases (represented as orange line) and the binding state increases (represented as green line).

References

Johari, Y. B., Scarrott, J. M., Pohle, T. H., Liu, P., Mayer, A., Brown, A. J., & James, D. C. (2022). Engineering of the CMV promoter for controlled expression of recombinant genes in HEK293 cells. Biotechnology Journal, 17(8), 2200062. Sequence and Features