Team:Lethbridge/Basic Part

iGEM

Basic Parts

Part BBa_K1791000

Part BBa_K1791000 is a theophylline ribozyme with an AU rich region flanking both sides of the part in order to aid proper functioning. When expressed in vivo, ribozyme cleavage can be induced by the addition of theophylline. This is not the case in vitro which results in cleavage of the ribozyme with or without the addition of theophylline. During design the ribozyme was further tested in vitro to prevent it from cleaving without the addition of theophylline; under varying temperatures, magnesium and spermidine concentrations. We were unable to prevent ribozyme cleavage without the addition of theophylline in vitro. We will be investigating mutations to the theophylline aptamer domain in order to optimize this part under in vitro conditions. However this part has been shown to work effectively under in vivo conditions. [1]

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References

  1. Win and Smolke (2007). A modular and extensible RNA-based gene-regulatory platform for engineering cellular function. PNAS. 104(36):14283-8.