Genomic Editing by Peptide Mediated Delivery of Crispr Endonucleases and Guide RNA’s

Gene therapy is a new medical discipline that offers a cure for patients who are born with genetic defects or suffer from progressive diseases such as cancer and neurodegeneration. The advantages it offers over drug therapy are attributed to its capability to target the underlying cause of these diseases and not just their symptoms. Cas9 and Cpf1 are endonucleases that have been recently identified in the Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR) bacterial immune system that hold great promise in gene therapy due to their high precision and efficiency in gene editing. Here, we aim to accomplish a transfection reagent-free delivery of CRISPR nucleases into stably fluorescent HEK293 cells using biocompatible cationic peptides in order to knock-out an endogenously expressed EGFP gene. The achieved expression and delivery of recombinant functional Cas9 protein using cationic peptides serves as a proof-of-concept for future gene editing and cell-specific targeting in different cell models in the future.