The Sendai virus vector has received a lot of attention due to its broad tropism for mammalian cells. As a result of efforts for genetic studies based on a mutant virus, we can now express more than 10 genes of up to 13.5 kilo nucleotides in a single vector with high protein expression efficiency. To prove this benefit, we examined the efficacy of the novel ribonucleic acid (RNA) virus vector harboring the human iduronate-2-sulfatase (IDS) gene with 1,653 base pairs, a causative gene for mucopolysaccharidosis type II, also known as a disorder of lysosomal storage disorders. As expected, this novel RNA vector with the human IDS gene exhibited its marked expression as determined by the expression of enhanced green fluorescent protein and IDS enzyme activity. While these cells exhibited a normal growth rate, the BHK-21 transformant cells stably expressing the human IDS gene persistently generated an active human IDS enzyme extracellularly. The human IDS protein produced failed to be incorporated into the lysosome when cells were pretreated with mannose-6-phosphate, demonstrating that this human IDS enzyme has potential for therapeutic use by cross-correction. These results suggest that our novel RNA vector may be applicable for further clinical settings.

中文翻译：

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用新型单链 RNA 病毒载体生产治疗性艾杜糖酸-2-硫酸酯酶

仙台病毒载体因其对哺乳动物细胞的广泛趋向性而备受关注。作为基于突变病毒的遗传研究努力的结果，我们现在可以在单个载体中以高蛋白质表达效率表达 10 多个高达 13.5 千核苷酸的基因。为了证明这种益处，我们检测了新型核糖核酸 (RNA) 病毒载体的功效，该病毒载体包含具有 1,653 个碱基对的人类艾杜糖酸 2-硫酸酯酶 ( IDS ) 基因，该基因是 II 型粘多糖贮积症的致病基因，也称为溶酶体贮积症。正如预期的那样，这种带有人类IDS 的新型 RNA 载体基因表现出其显着的表达，这是由增强的绿色荧光蛋白的表达和 IDS 酶活性所确定的。虽然这些细胞表现出正常的生长速度，但稳定表达人IDS基因的 BHK-21 转化细胞持续在细胞外产生活性人 IDS 酶。当用 6-磷酸甘露糖预处理细胞时，产生的人类 IDS 蛋白未能整合到溶酶体中，这表明这种人类 IDS 酶具有通过交叉校正进行治疗的潜力。这些结果表明我们的新型 RNA 载体可能适用于进一步的临床环境。