In your study, you suspect a Y protein which may play role in ER stress. Design an experiment to analyse role of Y protein (in vitro or in vivo) based on CRISPR/cas9 manipulation systems. Additionaly, how to identify expression localizations of this gene without any genomic alterations. In your experiments, what can you do to reduce off-target effect of CRISPR systems. (Hints: Do not forget to use control groups and specify necessary steps and analysis )