Receptor Mediated Delivery of Cas9-Nanobody Induces Cisplatin Synthetic Dose Sensitivity

Abstract

26The CRISPR/Cas9 system has shown great potential for precisely editing genomic DNA 27 sequences by introducing site-specific DNA cuts that are subsequently repaired by the cell. 28However, delivery of the CRISPR ribonucleoprotein remains an understudied area and hinders 29 realizing the full potential of the system. We prepared Cas9 ribonucleoprotein complexes 30 chemically conjugated to the 7D12 nanobody and demonstrate receptor-mediated transfection of 31Cas9 into A549 non-small-cell lung cancer cells via binding to the epithelial growth factor receptor 32 for subsequent cell internalization. We further show that transfection with a Cas9 33 ribonucleoprotein targeting the BRCA2 gene results in an enhanced sensitivity to the 34 chemotherapeutic drug Cisplatin, and thereby induces a synthetic dose lethality in A549 cells. 35 36