Coding

Part:BBa_K4586021

Designed by: Mohamed Mohamed Saad Aboelghare   Group: iGEM23_AFCM-Egypt   (2023-09-15)
Revision as of 15:35, 24 September 2023 by Ahmed Mattar (Talk | contribs)


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.

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


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]


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Categories
Parameters
None