Part:BBa_K4586016
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 Syn-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 Syn-Notch receptor and BCR.
Graph(1) shows an increase in the free portion of CCP1 of Syn-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).
Presence of the CMV part controles booster gene expression which increases the level of engineered exosomes so it plays an effective role to increase the efficacy of the therapeutic agent.
Booster gene with conditioned release
This Represents the relation between the activation of the internal domain of the Syn-Notch (represented as red line) and production of exosomes with specific cargo (represented as blue line) which increases in their level due to presence of the booster gene, Where the production of the engineered exosomes is initiated once the internal domain is activated.
No booster genes with conditioned release
This Represents the relation between the activation of the internal domain of the Syn-Notch (represented as red line) and production of exosomes with specific cargo (represented as blue line) as the production of the engineered exosomes is initiated once the internal domain is activated.
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
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]
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