Team:London Biohackspace/experiments/rna-based-regulation

Experiments

RNA INTERFERENCE BASED REGULATION OF RFP IN E. COLI

Introduction

To test whether paired-termini antisense RNA interference can be used to regulate gene expression a partial mRFP knockdown BioBrick will be synthesized and compared to the existing mRFP knockdown BioBrick created by Hokkaido Univeristy iGEM 2014 team. This part will contain a number of mismatched bases in the hairpin structure in an effort to reduce overall stability of the RNAi molecule and subsequently reduce its ability to knockdown the mRFP transcript. The RBS+mRFP (BBa_K516032) and RBS+mRFP RNA interference (BBa_K1524104) BioBricks created by the Hokkaido University 2014 iGEM team will be ligated to a constitutive promoter (BBa_J23103) found within the 2015 distribution kit. Our partial knockdown part will be ligated with the same promoter and each knockdown will be co-transformed with the mRFP expressing plasmid. RFP floresence will then be measured to determine whether the partial knockdown part we created reduces RFP expression when compared to the full knockdown part.

Materials and methods

Results

Discussion