Regulatory

Part:BBa_K3504004

Designed by: Ahmed Wael   Group: iGEM20_AFCM-Egypt   (2020-09-19)
Revision as of 09:27, 23 October 2020 by Ahmed Wael (Talk | contribs) (→‎Characterization)


ِِAlphavirus Subgenomic Promoter 30

Part Description

Alpha-virus replicon Subgenomic constitutive promoter. And a subgenomic promoter is a promoter added to a virus for a specific heterologous gene resulting in the formation of mRNA for that gene alone.

Usage

SG RNA is transcribed from the SG promoter and serves as template for translation of viral structure protein e.g.:capsid,E2 and E1 which gives alpha virus attractive systems for designing self replicating vectors for delivery and expression of heterogeneous genetic information. It’s also used for amplification and replication by the VEEV replication complex SG RNA is transcribed from a promoter located in the alpha virus specific RNA replication intermediate and is not further amplified. SG promoter transcribes RNA encoding proteins of interest and encodes the same (3 CSE ) and poly A tail as the viral genome but contains a different (5 UTR) and lacks the (51-nt CSE). We used it as a control to demonstrates a standard level of RNA synthesis and heterogeneous protein expression.

Characterization

We have made simulations using mathematical modelling techniques which showed expression values relative to those present in literature as shown in figures 1 & 2. These findings suggest that SGP 30 & 15 show the highest expression values with SGP 15 being the highest

Figure 1.This figure shows mathmatical modelling simulations illustrations expression variation between SGP30 vs SGP15 which both show high expression however SGP 15 show much higher expression .
Figure 2. shows mvenus expression in different variants of subgenomic promoters.

















References

Wagner, T. E., Becraft, J. R., Bodner, K., Teague, B., Zhang, X., Woo, A., ... & Sanders, N. N. (2018). Small-molecule-based regulation of RNA-delivered circuits in mammalian cells. Nature chemical biology, 14(11), 1043-1050. 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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