A Nanoparticle Platform for Accelerated In Vivo Oral Delivery Screening of Nucleic Acids

Abstract Oral administration of nucleic acids, such as small interfering RNAs and antisense oligonucleotides, represents a potent modality for treating many gastrointestinal (GI) diseases. However, their use is limited due to a lack of carriers that can effectively mediate their delivery through GI barriers, including gastric enzymes and mucus membranes. Lipid nanoparticles (LNPs) are an emerging delivery system that can protect nucleic acids from degradation and mediate their intracellular delivery, but few studies have evaluated their ability to overcome oral delivery barriers. Here, a high‐throughput in vivo screening platform is developed to accelerate the design of LNPs for oral delivery of nucleic acids. A library of LNPs that encapsulate DNA barcodes (b‐DNAs) was pooled and orally administered to mice, and their delivery was quantified by deep sequencing. Sequencing results indicated that increases in the molar ratio of cholesterol to lipid‐anchored polyethylene glycol conjugate in LNPs enhanced their GI content retention and that the presence of an LNP carrier improved b‐DNA delivery to GI tissues, relative to the delivery of naked b‐DNA. Collectively, these results suggest that high‐throughput in vivo screening can accelerate the discovery of LNPs for GI tract nucleic acid delivery upon oral administration.