Why- If the CRISPR-Cas9 transfection is not successful, it would not be relevant to relate the observations from transfected cells to the expected outcome of gene editing experiment.
How- CRISPR-Cas9’s successful transfection can be verified through its detection at the protein level by employing Western blot or confocal staining analysis of cells that were subjected to CRISPR-Cas9 transfection.
2. Localization of CRISPR-Cas9 at subcellular level
Why- CRISPR-Cas9 must translocate to the nuclei of the transfected cells for executing its nuclease activity on the genomic DNA. Therefore, one must check that the Cas9 protein is actually being delivered into the nucleus.
How - The...


