Department of Biomedical Engineering, Columbia University, New York, New York 10027, United States.
Biomacromolecules Research Team, RIKEN Center for Sustainable Resource Science, Saitama 351-0198, Japan.
Nano Lett. 2023 Feb 8;23(3):757-764. doi: 10.1021/acs.nanolett.2c02306. Epub 2023 Jan 17.
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.
有效递呈 CRISPR-Cas9 组件对于实现治疗潜力至关重要。尽管已经开发了许多用于该应用的递呈方法，但由于胃肠道的降解性质，尚未探索口服递呈。针对这个问题，我们开发了一系列新型苯硼酸（PBA）功能化壳聚糖-聚亚乙基亚胺（CS-PEI）聚合物用于口服 CRISPR 递呈。PBA 功能化使多聚物具有更高的稳定性、在黏液中更顺畅的传输以及在细胞内有效的内体逃逸和细胞质解包。从 12 种 PBA 功能化的 CS-PEI 多聚物文库中，我们确定了一种在口服灌胃后在小鼠肠道黏膜中显示出最有效渗透的配方。优化的配方表现出可行的 CRISPR 介导的靶蛋白下调和下游胆固醇减少。作为第一个口服 CRISPR 载体，这项研究表明了以患者依从的方式满足局部和全身编辑需求的潜力。
