In order to design a device that would produce a signal in response to the biomarker RNA, a special guide RNA was needed to bind to LwCas13a and miRNA424-5p. This guide RNA was designed using the crRNA pin-loop structure found in the Leptotrichia wadeii bacterium and combining it with the complement to miRNA424-5p.
These parts and our procedure will help other IGEM teams design their own guide RNAs and enable teams to incorporate Cas protein systems into future projects.
Our project contributed two new basic parts to the Registry that will enable future iGEM teams to quickly and easily perform their experiments. The function of miRNA424-5p is well documented, but it was not easily accessible as a BioBrick part. The crRNA that is used specifically by Cas13 is another part that will prove useful to teams that want to design their own Cas systems, whether for signaling or for removing specific genes in their projects. These parts are important because even if future iGEM teams don’t use our specific parts, knowing how our team used the parts can help inspire future projects.