Creation of novel cell-penetrating peptides for intracellular drug delivery using systematic phage display technology originated from tat transduction domain
Creation of novel cell-penetrating peptides for intracellular drug delivery using systematic phage display technology originated from tat transduction domain
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DOI：
10.1248/bpb.30.218
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发表时间：
2007-02-01
影响因子：
2
通讯作者：
Tsunoda, Shin-ichi
中科院分区：
文献类型：
--
作者：
Kamada, Haruhiko;Okamota, Takayuki;Tsunoda, Shin-ichi
许多生物活性蛋白质需要在细胞内递送以在细胞质内发挥其治疗作用。已经开发了细胞穿透肽（CPP），以将各种各样的货物以完全生物活性形式有效地递送到一系列细胞类型中，用于治疗多种临床前疾病模型。为了进一步发展这种方法，我们建立了一个系统的方法来确定新的CPP使用噬菌体展示技术。首先，我们从噬菌体肽库中筛选与细胞膜结合的肽。其次，为了评估作为细胞内载体的功能，我们将结合肽的cDNA与蛋白质合成抑制因子（PSIF）重组以产生融合蛋白。培养随机选择的克隆，诱导肽-PSIF融合蛋白的表达，然后筛选细胞中的蛋白质合成活性。使用这种系统的方法，新的和有效的CPP被迅速确定。我们认为，这些新的细胞穿透肽可用作蛋白质治疗的药物递送工具或研究蛋白质转导到细胞质中的机制的分析工具。
Many biologically active proteins need to be delivered intracellularly to exert their therapeutic action inside the cytoplasm. Cell penetrating peptides (CPPs) have been developed to efficiently deliver a wide variety of cargo in a fully biological active form into a range of cell types for the treatment of multiple preclinical disease models. To further develop this methodology, we established a systematic approach to identify novel CPPs using phage display technology. Firstly, we screened a phage peptide library for peptides that bound to the cell membrane. Secondly, to assess functionality as intracellular carriers, we recombined cDNAs of binding peptides with protein synthesis inhibitory factor (PSIF) to create fusion proteins. Randomly chosen clones were cultured and expression of peptide-PSIF fusion proteins induced, followed by screening of protein synthesis activity in cells. Using this systematic approach, novel and effective CPPs were rapidly identified. We suggest that these novel cell-penetrating peptides can utilized as drug delivery tools for protein therapy or an analytical tool to study mechanisms of protein transduction into the cytoplasm.