MOF effectively deliver CRISPR and enhance gene-editing efficiency via MOF’s hydrolytic activity of phosphate ester bonds

Abstract

Activity enhancement of gene-editing toolsets is critical not only to reverse the intractable endogenous mutagenesis, but also to develop new therapeutic modalities for clinical translation. However, the gene editing efficiency was often not fully satisfactory due to the deficient catalytic activity of CRISPR protein. Herein, we report a cocatalysis approach to improve gene editing efficiency using Zr6(μ3-O)4(μ3-OH)4-cluster-containing metal–organic framework (MOF), which can efficiently package Cpf1-encoding plasmids and deliver them into cellular cytosols. Importantly, Zr6(μ3-O)4(μ3-OH)4 cluster (building block of nanoPCN-8908) can pre-activate and hydrolyze the phosphate ester bonds (a type of key chemical bonds in between genomic DNAs needed to be cleaved before gene editing), assisting expressed Cpf1 to edit genomic loci. The editing efficiency of VEGFA and DNMT1 (two genes crucial to oncogenesis) by plasmid@nanoPCN-8908 is 5-fold and 7-fold, respectively, higher than did another MOF (ZIF-8) often used as the CRISPR vehicle. This MOF-based cocatalysis approach shows huge potential to promote the development of CRISPR gene editing technologies.