LAUSR

CRISPR/Cas9 has proven its accuracy and precision for gene editing by making a double-strand break at the predetermined nucleic acid sequence. Despite being a mainstream gene editing tool, CRISPR/Cas9 has limitations for its in vivo delivery due to the physico-chemical properties such as high molecular weight, supranegative charge, degradation in the presence of nucleases in the biological fluid, etc. Viral vector has been vastly used to deliver CRISPR/Cas components but possesses ample drawbacks and is challenging to translate. We hereby explored a cationic lipopolymer, i.e, mPEG b-(CB-{g-cationic chain; g-Chol; g-Morph}) for its efficiency in delivering CRISPR/Cas9 plasmid (pCas9) in vitro and in vivo. The polymer was utilized to form blank cationic nanoplexes having a zeta potential of +15.8 ± 0.7 mV. Being cationic, the blank nanoplexes were able to condense the pCas9 plasmid at a ratio of 1:20 with a complexation efficiency of ∼98% and showed a size and zeta potential of ∼141 ± 16 nm and 4.2 mV ± 0.7, respectively. The pCas9-loaded nanoplexes showed a transfection efficiency of ∼69% in ARPE-19 cells and showed ∼22% of indel frequency indicating the successful translation of Cas9 protein and guide RNA in the cytosol. Further, they were found to be stable under in vivo environment when given intravenously in Swiss albino mice. These lipopolymeric nanoplexes could be a potential carrier for CRISPR plasmids for genome editing applications. This article is protected by copyright. All rights reserved.