Stead Family Department of Pediatrics, Pappajohn Biomedical Institute, & Center for Gene Therapy, The University of Iowa, Iowa City, IA 52242, USA.
Biotechniques. 2020 May;68(5):235-239. doi: 10.2144/btn-2019-0150. Epub 2020 Mar 6.
Gene therapy for airway diseases requires efficient delivery of nucleic acids to the airways. In small animal models, gene delivery reagents are commonly delivered as a bolus dose. However, large animal models are often more relevant for the transition from preclinical studies to human trials. Aerosolizing viral vectors to the lungs of large animals can maximize anatomical distribution. Here, we describe a technique for aerosolization of viral vectors to the airways of newborn pigs. Briefly, a pig is anesthetized and intubated with an endotracheal tube, and a microsprayer is passed through the endotracheal tube. A fine mist is then sprayed into the distal trachea. Widespread and uniform distribution of transgene expression is critical for developing successful lung gene therapy treatments.
气道疾病的基因治疗需要将核酸有效递送到气道。在小动物模型中,基因传递试剂通常作为单次剂量给予。然而,大型动物模型通常与从临床前研究向人体试验的过渡更为相关。将病毒载体雾化到肺部可以最大限度地实现解剖分布。在这里,我们描述了一种将病毒载体雾化到新生猪气道的技术。简而言之,将猪麻醉并通过气管内管进行插管,然后将微喷雾器通过气管内管。然后将细雾喷入远端气管。广泛和均匀的转基因表达分布对于开发成功的肺部基因治疗至关重要。