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2012.igem.org:Wlc
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Project
We are designing a tool that will aid researchers in their study of stem cell differentiation, currently our project focuses on cardiomyocytes. We selected two promoters, one that is specific to atrial heart cells (of the gene ANF) and one that is specific to ventricular heart cells (of the gene MLC2v). The ANF promoter will drive the transcription of RFP, so cells of the atrial heart cell line will express RFP. The MLC2v promoter will drive the transcription of GFP, so cells of the ventricular heart cell line will express GFP.
Two plasmids will be synthesized. The first will contain an atrial cardiomyocyte specific promoter attached to GFP and puromycin resistance. The second will contain a ventricle cardiomyocyte specific promoter attached to RFP and neomycin. This allows us to select for stem cells that are expressing both plasmids by adding both neomycin and puromycin to our growth medium, so that all stem cells that do not contain the plasmids will die off. The formation of the stem cell line will allow researchers to study the differentiation pathway between Ventricular Cardiomyocytes and Atrial Cardiomyocytes by analyzing a change in the ratio of ventricular and atrial cardiomycites; which will be differentiable by the RFP and GFP expression for the particular cell
