...a template. In general, edits must be downstream (3’) of the nick site, and guides that target closer ...control of gene repression and activation. Cell , 159 (3), 647–661. https://doi.org/10.1016/j.cell.2014.09.029...Achieving your desired edit comes down to location, location, location. Positioning your chosen Cas enzyme ...designing the gRNA component for each category. Genetic Manipulation Application Cas Enzyme Additional Considerations...the table below, we highlight the most common categories of genetic manipulations used in mammalian cells...gene you target will depend on your specific application. Knockout : Target constitutively expressed regions...start sites that could allow for expression of a truncated protein. Targeting an exon coding for an essential...