Yin HaiFang, Moulton Hong, Betts Corinne, Wood Matthew
Anatomy and Genetics, Department of Physiology, University of Oxford, South Parks Road, Oxford, UK.
Methods Mol Biol. 2011;683:321-38. doi: 10.1007/978-1-60761-919-2_23.
Antisense oligonucleotides (AOs) are effective splice switching agents and have potential as therapeutics via the exclusion or inclusion of specific target gene exons to ameliorate and modify disease progression. The leading example is Duchenne muscular dystrophy (DMD), a fatal muscle degenerative disease, where AO-mediated skipping of specific DMD gene exons can restore the disrupted DMD open reading frame, leading to the production of functional dystrophin protein and ameliorate the DMD phenotype in animal models. Clinical proof-of-concept has recently been shown in two successful, independent Phase I clinical trials. These trials both followed local intramuscular treatments, and the challenge now is to develop and test systemic protocols, which will be required for treatment-aimed disease modification. Recently, a number of groups have demonstrated the promise of AOs directly conjugated to cell-penetrating peptides (CPPs) as having significant potential for systemic delivery and therapeutic correction in DMD animal models. Here, we review the background to this work and describe in detail the experimental protocols used in studies aimed at investigating CPP-conjugated AOs as systemic splice correcting agents in animal models of DMD.
反义寡核苷酸（AO）是有效的剪接转换剂，通过排除或包含特定靶基因外显子来改善和改变疾病进展，具有作为治疗药物的潜力。最典型的例子是杜氏肌营养不良症（DMD），这是一种致命的肌肉退行性疾病，在动物模型中，AO介导的特定DMD基因外显子跳跃可恢复中断的DMD开放阅读框，从而产生功能性肌营养不良蛋白，并改善DMD表型。最近在两项成功的独立I期临床试验中展示了临床概念验证。这两项试验均采用局部肌肉注射治疗，目前的挑战是开发和测试系统性方案，这对于旨在改变疾病的治疗是必需的。最近，一些研究小组已证明，直接与细胞穿透肽（CPP）偶联的AO在DMD动物模型中具有显著的全身递送和治疗矫正潜力。在此，我们回顾这项工作的背景，并详细描述旨在研究CPP偶联AO作为DMD动物模型中全身剪接校正剂的研究中所使用的实验方案。
