Difference between revisions of "Part:BBa K3504004"

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ِِAlphavirus Subgenomic Promoter 30

NOTICE: Parts in our range for this season have been created as a part of our Phase I design of our project. These parts HAVE NOT been tested or characterized in the lab due to COVID-19-related precautionary measures. We have enriched our new parts pages with data from literature and results from our modeling and simulations. If you are intending on using this part or others in our range, please keep in mind these limitations and update these parts with data from your experimentation. Feel free to reach us at: igem.afcm@gmail.com for further inquiries.

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