siRNA转染的增强由脂肪酸长度的优化和组氨酸在CPP内容<a href="#fn1" title="提供电子补充信息（ESI）。参见DOI：10.1039 / c9bm00688e">†</a>,Biomaterials Science

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siRNA转染的增强由脂肪酸长度的优化和组氨酸在CPP内容 †
Biomaterials Science ( IF 5.8 ) Pub Date : 2019-08-14 00:00:00 , DOI: 10.1039/c9bm00688e
Ly Porosk _{1,

2,

3,

4} , Piret Arukuusk _{1,

2,

3,

4} , Kaisa Põhako _{1,

2,

3,

4} , Kaido Kurrikoff _{1,

2,

3,

4} , Kristina Kiisholts _{1,

2,

3,

4} , Kärt Padari _{2,

3,

4,

5} , Margus Pooga _{1,

2,

3,

4} , Ülo Langel _{1,

2,

3,

4,

6}

Affiliation

细胞外合成核酸，例如siRNA，无法有效地达到其预期目标。因此，诸如细胞穿透肽（CPP）的转运方法增加了转运，可以增强siRNA作为治疗剂的效力。CPP NickFect55（NF55）是一种有效的基于肽的递送载体，先前已用于将质粒DNA体内递送至细胞。为了从所运送的货物中实现更高的细胞内递送和生物活性，我们通过优化CPP NF55序列中的pH敏感性，净电荷，疏水性和电荷分布，设计了一系列含组氨酸的肽。在当前的工作中，我们应用了一种策略，其中我们替换了肽C端的氨基酸，以便将疏水性和亲水性氨基酸分布到沿α螺旋投影的不同区域，包括组氨酸到序列中在N末端进行修饰，并优化N末端脂肪酸修饰以适合特定的肽序列。我们根据转染效率，CPP / siRNA复合物的稳定性以及CPP的特性（例如溶血活性，缓冲能力和细胞毒性）对CPP进行了测试。因此，我们推出了一种具有完全重新设计的N末端的新肽，可显示对其物理环境的适应性反应。这种肽– NickFect70（NF70）–有效地浓缩siRNA，保护货物免于降解，并有效介导哺乳动物细胞培养中靶基因的敲低和体内，在小鼠模型中。

"点击查看英文标题和摘要"

Enhancement of siRNA transfection by the optimization of fatty acid length and histidine content in the CPP†

Extracellular synthetic nucleic acids, such as siRNAs, are unable to reach their intended targets efficiently. Therefore, delivery methods such as cell-penetrating peptides (CPP), which increase their transport, could enhance the potency of siRNA as therapeutic agents. The CPP NickFect55 (NF55) is an efficient peptide-based delivery vector, which has been previously used to deliver plasmid DNA into cells in vivo. To achieve higher intracellular delivery and bioactivity from the delivered cargo, we designed a series of histidine-containing peptides by optimizing pH-sensitivity, net charge, hydrophobicity, and charge distribution in the sequence of the CPP NF55. In the current work, we have applied a strategy where we have replaced amino acids in the C-terminus of the peptide in order to distribute hydrophobic and hydrophilic amino acids into distinct regions along the alpha-helical projection, including histidine amino acids into the sequence at the N-terminus, and optimizing the N-terminal fatty acid modification to suit the specific peptide sequence. We tested the CPPs based on the transfection efficacy, CPP/siRNA complex stability, and the properties of the CPPs, such as hemolytic activity, buffering capability and cell toxicity. As a result, we have introduced a new peptide with a completely redesigned N-terminus that displays adaptive response to its physical environment. This peptide – NickFect70 (NF70) – efficiently condenses siRNA, protects the cargo against degradation and effectively mediates target gene knockdown both in mammalian cell culture and in vivo, in a mouse model.

更新日期：2019-08-14

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