Part:BBa_K1994021:Experience

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Applications of BBa_K1994021

This part was characterised by the Warwick iGEM 2016 team.

This part was also triple transformed into DH5alpha cells also containing pSB3T5-PP7-omega (BBa_K1994024) and a GFP-dCas9 construct containing a weakened omega promoter. 20 different targeting regions were cloned into the golden gate site of this sgRNA part. This was to demonstrate that the library of sgRNAs would increase the expression of GFP by using dCas9 binding to bring an omega factor closer to the weakened omega promoter. However the data collected shows negligible difference in GFP expression between cells containing the library sgRNAs and the negative control that contains all three plasmids, but without an inserted targeting region.

The white diamonds show the negative control with error bars representing a single standard deviation from the mean. The majority of the library sgRNAs fall within the error bars. The difference in expression is not sufficiently different to suggest a correlation between specific sgRNAs and GFP expression compared to the negative control.

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