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To create the Gemini parts library we made use of two “classes” of plasmid: guide RNA expressing vectors and guide RNA operator reporter vectors. When these two plasmids are co-transfected with a dCas9-VPR plasmid into a mammalian cell line, the production of a gene of interest proceeds. The genes of interest in our work were fluorescent proteins; specifically GFP was utilized in the guide RNA operator reporter vectors submitted to the registry. The guide RNA operator vector can be considered a composite part. It is constructed by combining the following parts in order: BBa_K1875009 (the target sequence), BBa_K1875000 (the miniCMV), BBa_K1875001 (the Kozak sequence), BBa_K1875003 (the GFP), BBa_K1875002 (the Rabbit Beta Globin PolyA). The sources of these parts can be found on their respective registry pages. We have submitted BBa_K1875018 for best composite part due to both its high expression and consistency in experimentation. Specifically, this part, when compared with a CMV, can express GFP at a high level than the strong promoter. This means that this part grants a scientist the ability to produce a protein at a higher level than a strong promoter and grant them a temporal control. | To create the Gemini parts library we made use of two “classes” of plasmid: guide RNA expressing vectors and guide RNA operator reporter vectors. When these two plasmids are co-transfected with a dCas9-VPR plasmid into a mammalian cell line, the production of a gene of interest proceeds. The genes of interest in our work were fluorescent proteins; specifically GFP was utilized in the guide RNA operator reporter vectors submitted to the registry. The guide RNA operator vector can be considered a composite part. It is constructed by combining the following parts in order: BBa_K1875009 (the target sequence), BBa_K1875000 (the miniCMV), BBa_K1875001 (the Kozak sequence), BBa_K1875003 (the GFP), BBa_K1875002 (the Rabbit Beta Globin PolyA). The sources of these parts can be found on their respective registry pages. We have submitted BBa_K1875018 for best composite part due to both its high expression and consistency in experimentation. Specifically, this part, when compared with a CMV, can express GFP at a high level than the strong promoter. This means that this part grants a scientist the ability to produce a protein at a higher level than a strong promoter and grant them a temporal control. | ||
Revision as of 19:50, 15 October 2016
To create the Gemini parts library we made use of two “classes” of plasmid: guide RNA expressing vectors and guide RNA operator reporter vectors. When these two plasmids are co-transfected with a dCas9-VPR plasmid into a mammalian cell line, the production of a gene of interest proceeds. The genes of interest in our work were fluorescent proteins; specifically GFP was utilized in the guide RNA operator reporter vectors submitted to the registry. The guide RNA operator vector can be considered a composite part. It is constructed by combining the following parts in order: BBa_K1875009 (the target sequence), BBa_K1875000 (the miniCMV), BBa_K1875001 (the Kozak sequence), BBa_K1875003 (the GFP), BBa_K1875002 (the Rabbit Beta Globin PolyA). The sources of these parts can be found on their respective registry pages. We have submitted BBa_K1875018 for best composite part due to both its high expression and consistency in experimentation. Specifically, this part, when compared with a CMV, can express GFP at a high level than the strong promoter. This means that this part grants a scientist the ability to produce a protein at a higher level than a strong promoter and grant them a temporal control.