Data from: Lung and liver editing using lipid nanoparticle delivery of a stable CRISPR-Cas9 RNP

Lipid nanoparticle (LNP) delivery of CRISPR ribonucleoproteins (RNPs)
could enable high-efficiency, low-toxicity, and scalable in vivo genome
editing if efficacious RNP:LNP complexes can be reliably produced. Here,
we engineered a thermostable Cas9 from Geobacillus stearothermophilus
(GeoCas9) to generate iGeoCas9 variants capable of >100X more
genome editing of cells and organs compared to the native GeoCas9 enzyme.
Furthermore, iGeoCas9 RNP:LNP complexes edited a variety of cell types and
induced homology-directed repair (HDR) in cells receiving co-delivered
single-stranded DNA (ssDNA) templates. Using tissue-selective LNP
formulations, we observed genome editing levels of 16‒37% efficiency in
the liver and lungs of model reporter mice that received single
intravenous injections of iGeoCas9 RNP:LNPs. In addition, iGeoCas9 RNP
complexed to biodegradable LNPs edited the disease-causing SFTPC gene in
lung tissue with 19% average efficiency, representing a major improvement
over genome editing levels observed previously using viral or non-viral
delivery strategies. These results show that thermostable Cas9 RNP:LNP
complexes are a powerful alternative to mRNA:LNP delivery vehicles and
expand the therapeutic potential of genome editing.