Difference between revisions of "Part:BBa K4586021"

Revision as of 15:53, 5 October 2023

VP64

Part Description

This parts codes for VP64 which is transcriptional activator peptide formed of two components, first four tandem repeats of VP16 (Herpes simplex viral protein) second, amino acids 437-447*. This transcription module is a classic molecular biology tool due to its strong transcriptional activity that can be used for conditional gene expression by fusing to it another protein domain as ZF peptide that can bind near the promoter of the gene of interest.

Usage

We implemented this part in our design to trigger the transcriptional activity of ZF21.61 minCMV promoter that control the expression of our therapeutic agent or cargo (Cas12k). Therefore, VP64 share in forming the composite part ZF21.16-VP64 that represent the internal domain of our syn notch receptor which design to be activated after cell to cell interaction between MSC and B-cell receptor as shown in figure 1.

Figure 1: This figure illustrates the design of our biological circuit expressing our therapeutic agent under the control of the VP64 transcription module that regulates the activity of the ZF21.16minCMV promoter.

Literature Characterization

vp64 were transfected into HEK293T reporter cells and treated with abscisic acid or DMSO for 48 h.

Statistical significance was calculated with one-way ANOVA, using Dunnett’s multiple testing correction which showed that GFP expression was higher with VP64 than with any other promoter.

charactrization by mathematical modeling

This internal domain is activated after cell to cell interaction between MSC and B-cell receptor. That leads to increase of transcription factor VP64 that triggers the expression of the internal circuit that secretes the exosome's cargo then cargo loading to exosomes through the loading system (CD63-L7Ae).

As the expression of the internal domain increases (represented as red line), the transcription factor VP64 increases, increasing the expression of the internal circuit for the exosome's cargo to finally produce modified exosomes (represented in blue line).

References

Alerasool, N., Leng, H., Lin, Z. Y., Gingras, A. C., & Taipale, M. (2022). Identification and functional characterization of transcriptional activators in human cells. Molecular cell, 82(3), 677-695. Sequence and Features