What is the advantage of having an mCherry reporter gene in the sgRNA vector?
What is the advantage of having an mCherry reporter gene in the sgRNA vector?
The red fluorescent protein mCherry is expressed from the same vector that expresses each sgRNA and is included to simplify the process of determining the optimal MOI for the screen. The aim is to obtain a transduction efficiency of 30–40% in your target cells, which ensures that most transduced cells contain only a single sgRNA construct. For the calculation of transduction efficiency, a small-scale titration is performed and cells are analyzed for mCherry expression by flow cytometry or microscopy.