A Single Administration of CRISPR/Cas9 Lipid Nanoparticles Achieves Robust and Persistent In Vivo Genome Editing

• Published in: Cell reports (Cambridge) Vol. 22; no. 9; pp. 2227 - 2235
• Main Authors: Finn, Jonathan D., Smith, Amy Rhoden, Patel, Mihir C., Shaw, Lucinda, Youniss, Madeleine R., van Heteren, Jane, Dirstine, Tanner, Ciullo, Corey, Lescarbeau, Reynald, Seitzer, Jessica, Shah, Ruchi R., Shah, Aalok, Ling, Dandan, Growe, Jacqueline, Pink, Melissa, Rohde, Ellen, Wood, Kristy M., Salomon, William E., Harrington, William F., Dombrowski, Christian, Strapps, Walter R., Chang, Yong, Morrissey, David V.
• Format: Journal Article
• Language: English
• Published: United States Elsevier Inc 27.02.2018; Elsevier
• Subjects: Animals; Base Sequence; Cas9; CRISPR; CRISPR-Associated Protein 9 - metabolism; CRISPR-Cas Systems - genetics; CRISPR/Cas9; Gene Editing; gene therapy; Gene Transfer Techniques; genome editing; lipid nanoparticle; Lipids - chemistry; Liver - metabolism; liver delivery; LNP; Mice; Nanoparticles - administration & dosage; Nanoparticles - chemistry; Rats; RNA, Guide, CRISPR-Cas Systems - chemistry; RNA, Guide, CRISPR-Cas Systems - genetics; sgRNA; TTR; CRISPR; genome editing; lipid nanoparticle; TTR; CRISPR/Cas9; sgRNA; gene therapy; Cas9; liver delivery; LNP
• ISSN: 2211-1247; 2211-1247
• DOI: 10.1016/j.celrep.2018.02.014

The development of clinically viable delivery methods presents one of the greatest challenges in the therapeutic application of CRISPR/Cas9 mediated genome editing. Here, we report the development of a lipid nanoparticle (LNP)-mediated delivery system that, with a single administration, enabled significant editing of the mouse transthyretin (Ttr) gene in the liver, with a >97% reduction in serum protein levels that persisted for at least 12 months. These results were achieved with an LNP delivery system that was biodegradable and well tolerated. The LNP delivery system was combined with a sgRNA having a chemical modification pattern that was important for high levels of in vivo activity. The formulation was similarly effective in a rat model. Our work demonstrates that this LNP system can deliver CRISPR/Cas9 components to achieve clinically relevant levels of in vivo genome editing with a concomitant reduction of TTR serum protein, highlighting the potential of this system as an effective genome editing platform. [Display omitted] •LNP delivery achieves >97% target protein knockdown for at least 12 months•Editing level is cumulative following multiple LNP doses•A sgRNA chemical modification pattern was critical for high levels of in vivo activity•Biodegradable lipid and CRISPR/Cas9 components are transient and well tolerated Finn et al. describe the development of a transient, biodegradable LNP-based CRISPR/Cas9 delivery system that achieves >97% knockdown of serum TTR levels following a single administration. Editing levels were stable for 12 months, despite the transient nature of the delivery system and the editing components.