partial functionality (Hug et al. 2016). In order to obtain a complete knockout allele, it is therefore optimal to target a constitutive exon at least 30–40 amino acids into the protein coding sequence that is not in the final exon. Even in this situation, a complete knockout cannot be guaranteed, since NMD does not always act efficiently, and aberrant splicing can occur to reconstitute at least part of the protein function. Thus, analysis of protein expression is an important aspect to consider with such mutations.
