A facile nonviral method for delivering genes and siRNAs to skeletal muscle of mammalian limbs
A facile nonviral method for delivering genes and siRNAs to skeletal muscle of mammalian limbs
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DOI：
10.1016/j.ymthe.2004.05.004
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发表时间：
2004-08-01
影响因子：
12.4
通讯作者：
Wolff, JA
中科院分区：
文献类型：
--
作者：
Hagstrom, JE;Hegge, J;Wolff, JA
递送越来越被认为是阻碍基于核酸的疗法（包括基因疗法和最近的基于sirna的疗法）巨大前景的关键障碍。虽然已经确定了许多具有治疗潜力的候选基因（和siRNA序列），但由于低效和/或不安全的递送技术，它们的效用尚未实现。我们现在描述了一种血管内、非病毒的方法，使核酸能够有效和可重复地传递到哺乳动物肢体肌肉的肌肉细胞（肌纤维）。该过程包括将裸露的质粒DNA或siRNA注射到肢体的远端静脉，该远端静脉被止血带或血压袖带暂时隔离。核酸以足够的体积快速注射，使核酸溶液外渗到肌肉组织中，从而促进了向肌纤维的递送。在小型和大型动物的骨骼肌中实现了高水平的转基因表达，毒性很小。也获得了siRNA传递到肢体肌肉的证据。该方法简单、有效、安全，适用于肢体肌肉基因治疗。
Delivery is increasingly being recognized as the critical hurdle holding back the tremendous promise of nucleic acid-based therapies that include gene therapy and more recently siRNA-based therapeutics. While numerous candidate genes (and siRNA sequences) with therapeutic potential have been identified, their utility has not yet been realized because of inefficient and/ or unsafe delivery technologies. We now describe an intravascular, nonviral methodology that enables efficient and repeatable delivery of nucleic acids to muscle cells (myofibers) throughout the limb muscles of mammals. The procedure involves the injection of naked plasmid DNA or siRNA into a distal vein of a limb that is transiently isolated by a tourniquet or blood pressure cuff. Nucleic acid delivery to myofibers is facilitated by its rapid injection in sufficient volume to enable extravasation of the nucleic acid solution into muscle tissue. High levels of transgene expression in skeletal muscle were achieved in both small and large animals with minimal toxicity. Evidence of siRNA delivery to limb muscle was also obtained. The simplicity, effectiveness, and safety of the procedure make this methodology well suited to limb muscle gene therapy applications.