Delivering functional gene into targeted skin cells or tissues to modulate the genes expression, has the potential to treat various hereditary cutaneous disorders. Nevertheless, the lack of safe and effective gene delivery vehicles greatly limits the clinical translation of gene therapy for inherited skin diseases. Herein, we developed a facile elution fractionation strategy to isolate eight HPAEs with ranging from 7.6 to 131.8 kg/mol and < 2.0 from the one crude HPAE, and investigated the expression efficiency for and plasmids. Gene transfection results revealed that the intermediate MW HPAEs, HPAE, exhibited the highest gene transfection efficiency (46.4%) and the strongest mean fluorescence intensity (143,032 RLU), compared to other isolated components and the crude product. Importantly, best-performing isolated HPAE effectively delivered (15,974 bp) and (7181 bp) plasmids, promoting the efficient expression of type VII collagen (C7) and transglutaminase-1 proteins in cutaneous cells. Our study establishes a straightforward step-by-step elution fractionation strategy for the development of HPAEs gene delivery vectors, expediting their clinical translation in inherited skin diseases.

中文翻译：

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通过逐步分级分离，高度支化的聚（β-氨基酯）介导难以转染的皮肤细胞中 COL7A1 和 TGM1 基因的出色表达

将功能基因传递到目标皮肤细胞或组织中以调节基因表达，有可能治疗各种遗传性皮肤病。然而，缺乏安全有效的基因递送载体极大地限制了遗传性皮肤病基因治疗的临床转化。在此，我们开发了一种简便的洗脱分级策略，从一种粗 HPAE 中分离出 8 种 HPAE，其浓度范围为 7.6 至 131.8 kg/mol，且 < 2.0，并研究了 和 质粒的表达效率。基因转染结果表明，与其他分离组分和粗产物相比，中间分子量的HPAE（HPAE）表现出最高的基因转染效率（46.4％）和最强的平均荧光强度（143,032 RLU）。重要的是，性能最佳的分离 HPAE 有效递送 (15,974 bp) 和 (7181 bp) 质粒，促进皮肤细胞中 VII 型胶原 (C7) 和转谷氨酰胺酶-1 蛋白的有效表达。我们的研究建立了一种简单的逐步洗脱分级策略，用于开发 HPAE 基因传递载体，加快其在遗传性皮肤病中的临床转化。