Team:WPI Worcester/Part Collection

CRISPR/dCas9 Editing System and Reporter Parts

The team created a total of 16 new BioBrick parts for submission to the registry. Each of these parts were either part of the editing system itself or were used to test the editing system. The core of the editing system made up several parts in the collection. Each contained the dCas9/editing enzyme fusion. The two editing enzymes used were ADAR1 and APOBEC1. Each dCas9/editing enzyme fusion had three different varients. Each varient had different lengths of spacer sequence called XTEN Linkers. XTEN1,XTEN2, and XTEN3 linkers were used for each editing enzyme The other parts of the editing system included the guide RNA which was on the same plasmid as the gene for the RFP protein mCherry. Finally, within this collection is the different reporter plasmids used to test the editing systems. For the APOBEC1 enzyme, an eGFP molecule with a 5' untranslated region and an ACG start codon was created. Additionally, a "wild type" eGFP molecule was created with it that contained a 5' untranslated region and an ATG

The reporters can be found here, Wild Type GFP,Mutated Start Codon GFP,Premature Termination Codon Globin with GFP, Wild Type Globin with GFP.

The editors can be found here, APOBEC 1XTEN,APOBEC 2XTEN,APOBEC 3XTEN, ADAR1 1XTENADAR1 2XTENADAR1 3XTENADAR2 1XTENADAR2 2XTENADAR2 3XTEN