@article{c1ae7dbfe6854ca7bd8d769feee680f5,
title = "Phenylboronic Acid-Functionalized Polyplexes Tailored to Oral CRISPR Delivery",
abstract = "Effective delivery of the CRISPR-Cas9 components is crucial to realizing the therapeutic potential. Although many delivery approaches have been developed for this application, oral delivery has not been explored due to the degradative nature of the gastrointestinal tract. For this issue, we developed a series of novel phenylboronic acid (PBA)-functionalized chitosan-polyethylenimine (CS-PEI) polymers for oral CRISPR delivery. PBA functionalization equipped the polyplex with higher stability, smooth transport across the mucus, and efficient endosomal escape and cytosolic unpackaging in the cells. From a library of 12 PBA-functionalized CS-PEI polyplexes, we identified a formulation that showed the most effective penetration in the intestinal mucosa after oral gavage to mice. The optimized formulation performed feasible CRISPR-mediated downregulation of the target protein and reduction in the downstream cholesterol. As the first oral CRISPR carrier, this study suggests the potential of addressing the needs of both local and systemic editing in a patient-compliant manner.",
keywords = "CRISPR-Cas9, Chitosan, Genome editing, Oral gene delivery, Phenylboronic acid",
author = "Naoto Yoshinaga and Zhou, {Joyce K.} and Cong Xu and Quek, {Chai Hoon} and Yuefei Zhu and Ding Tang and Hung, {Lin Yung} and Najjar, {Sarah A.} and Shiu, {Chin Ying Angela} and Margolis, {Kara Gross} and Lao, {Yeh Hsing} and Leong, {Kam W.}",
note = "Funding Information: We would like to acknowledge the technical support from the Flow Cytometry Core Facility and Confocal and Specialized Microscopy Shared Resource of Herbert Irving Comprehensive Cancer Center at Columbia University, funded in part through NIH Center Grant (P30CA013696). Lightsheet imaging was carried out under the assistance of Dr. Humberto Avila in the Cellular Imaging Core Facility at Columbia-Zuckerman Institute, funded through NIH Instrumentation Program (1S10OD023587-01). The animal study was approved by the Institutional Animal Care and Use Committees at Columbia University (AC-AABD5600, AC-AAAZ4486). Figure 2 and TOC were created using BioRender. Funding Information: This work was supported by NIH (K.W.L., UG3-NS115598) and the Career Development Award from American Society of Gene & Cell Therapy (Y.-H.L., ASGCT2021003). It was also partially supported by DARPA (K.W.L., HR00111920009) on the synthesis and characterization of the polymeric gene carriers. N.Y. acknowledges the support of the JSPS Overseas Research Fellowship from Japan Society for the Promotion of Science. The content is solely the responsibility of the authors and does not necessarily represent the official views of the funders. Funding Information: We would like to acknowledge the technical support from the Flow Cytometry Core Facility and Confocal and Specialized Microscopy Shared Resource of Herbert Irving Comprehensive Cancer Center at Columbia University, funded in part through NIH Center Grant (P30CA013696). Lightsheet imaging was carried out under the assistance of Dr. Humberto Avila in the Cellular Imaging Core Facility at Columbia-Zuckerman Institute, funded through NIH Instrumentation Program (1S10OD023587-01). The animal study was approved by the Institutional Animal Care and Use Committees at Columbia University (AC-AABD5600, AC-AAAZ4486). and TOC were created using BioRender. Publisher Copyright: {\textcopyright} 2023 American Chemical Society.",
year = "2023",
month = feb,
day = "8",
doi = "10.1021/acs.nanolett.2c02306",
language = "English (US)",
volume = "23",
pages = "757--764",
journal = "Nano Letters",
issn = "1530-6984",
publisher = "American Chemical Society",
number = "3",
}