The ability of cell-penetrating peptides to cross plasma membranes has been used for various applications, including the delivery of bioactive molecules to inhibit disease-producing cellular mechanisms. Selective drug delivery into target cells improves drug distribution and decreases dosing and toxicity. In this review, the authors outline the main challenges in the field, namely clarification of mechanisms of entry into cells, as well as current and future perspectives regarding cell-penetrating peptides application for human therapeutics. Here, the authors discuss some of the factors that influence efficacy of delivery and review the current status of preclinical studies and clinical trials involving the use of cell-penetrating peptide-mediated delivery of therapeutics.