Cell-Penetrating Peptide-Mediated Delivery of CRISPR-Cas9

Currently, the delivery of the two components of CRISPR-Cas9 system has been conducted using either plasmid or viral vectors or the direct delivery of the protein and RNA into the cells, with direct delivery boasting several advantages over the plasmid approach.

Direct delivery results in shorter exposure time at the cellular level, which in turn leads to lower toxicity and fewer off-target mutations. Here, a cell-penetrating peptide (CPP), with the ability to translocate across cell membranes, has been adopted as a way of efficient delivery of the Cas9 protein and guide RNA.

The team of Bharathi Suresh from Yonsei University in South Korea has developed a method for treating human cell lines with CPP-conjugated recombinant Cas9 protein and CPP-complexed guide RNAs. Their study also describes a protocol for preparing the CPP-conjugated recombinant Cas9 protein and CPP-complex guide RNAs, as well as methods to achieve safe genome editing in human cell lines.

For more on this study, read the article in Methods in Molecular Biology.