Fast and Efficient CRISPR/Cas9 Genome Editing In Vivo Enabled by Bioreducible Lipid and Messenger RNA Nanoparticles

• Published in: Advanced materials (Weinheim) Vol. 31; no. 33; pp. e1902575 - n/a
• Main Authors: Liu, Ji, Chang, Jin, Jiang, Ying, Meng, Xiandi, Sun, Tianmeng, Mao, Lanqun, Xu, Qiaobing, Wang, Ming
• Format: Journal Article
• Language: English
• Published: Germany Wiley Subscription Services, Inc 01.08.2019
• Subjects: Animals; Biological Transport; Biomedical materials; Cell Line, Tumor; CRISPR; CRISPR-Associated Protein 9 - genetics; CRISPR/Cas9; Editing; Fluorescence; Gene Editing - methods; Gene Knockdown Techniques - methods; Gene Transfer Techniques; genome editing; Genomes; Green Fluorescent Proteins - genetics; Green Fluorescent Proteins - metabolism; Humans; lipid nanoparticles; Lipids; Lipids - chemistry; Materials science; messenger RNA delivery; Mice; Nanoparticles; Nanoparticles - chemistry; Nuclease; Oxidation-Reduction; Proprotein Convertase 9 - genetics; Proprotein Convertase 9 - metabolism; Proteins; RNA, Guide, CRISPR-Cas Systems - administration & dosage; RNA, Guide, CRISPR-Cas Systems - chemistry; RNA, Messenger - administration & dosage; RNA, Messenger - chemistry; genome editing; lipid nanoparticles; messenger RNA delivery; CRISPR/Cas9
• DOI: 10.1002/adma.201902575
• ISSN: 0935-9648; 1521-4095

A main challenge to broaden the biomedical application of CRISPR/Cas9 (clustered regularly interspaced short palindromic repeat (CRISPR) associated protein 9) genome editing technique is the delivery of Cas9 nuclease and single‐guide RNA (sgRNA) into the specific cell and organ. An effective and very fast CRISPR/Cas9 genome editing in vitro and in vivo enabled by bioreducible lipid/Cas9 messenger RNA (mRNA) nanoparticle is reported. BAMEA‐O16B, a lipid nanoparticle integrated with disulfide bonds, can efficiently deliver Cas9 mRNA and sgRNA into cells while releasing RNA in response to the reductive intracellular environment for genome editing as fast as 24 h post mRNA delivery. It is demonstrated that the simultaneous delivery of Cas9 mRNA and sgRNA using BAMEA‐O16B knocks out green fluorescent protein (GFP) expression of human embryonic kidney cells with efficiency up to 90%. Moreover, the intravenous injection of BAMEA‐O16B/Cas9 mRNA/sgRNA nanoparticle effectively accumulates in hepatocytes, and knocks down proprotein convertase subtilisin/kexin type 9 level in mouse serum down to 20% of nontreatment. The leading lipid nanoparticle, BAMEA‐O16B, represents one of the most efficient CRISPR/Cas9 delivery nanocarriers reported so far, and it can broaden the therapeutic promise of mRNA and CRISPR/Cas9 technique further. A bioreducible lipid nanoparticle integrated with disulfide bonds can efficiently deliver Cas9 messenger RNA (mRNA) and single‐guide RNA into cells, while releasing mRNA in response to the reductive intracellular environment for genome editing as fast as 24 h post mRNA delivery. The leading lipid nanoparticle, BAMEA‐O16B, represents one of the most efficient CRISPR/Cas9 delivery nanocarriers reported so far.