内化
内体
细胞生物学
绿色荧光蛋白
细胞
化学
蛋白质片段互补分析
计算生物学
互补
转导(生物物理学)
细胞培养
细胞质
生物
生物化学
基因
表型
遗传学
作者
Nadia Milech,Brooke Allison Cattell Longville,Paula Cunningham,Marie N. Scobie,Heique Marlis Bogdawa,Scott Winslow,Mark Anastasas,Theresa Connor,Ferrer Ong,Shane R. Stone,Maria Kerfoot,Tatjana Heinrich,Karen M. Kroeger,Yew‐Foon Tan,Katrin Hoffmann,Wayne R. Thomas,Paul M. Watt,R.M. Hopkins
摘要
Abstract Efficient cargo uptake is essential for cell-penetrating peptide (CPP) therapeutics, which deliver widely diverse cargoes by exploiting natural cell processes to penetrate the cell’s membranes. Yet most current CPP activity assays are hampered by limitations in assessing uptake, including confounding effects of conjugated fluorophores or ligands, indirect read-outs requiring secondary processing and difficulty in discriminating internalization from endosomally trapped cargo. Split-complementation Endosomal Escape (SEE) provides the first direct assay visualizing true cytoplasmic-delivery of proteins at biologically relevant concentrations. The SEE assay has minimal background, is amenable to high-throughput processes and adaptable to different transient and stable cell lines. This split-GFP-based platform can be useful to study transduction mechanisms, cellular imaging and characterizing novel CPPs as pharmaceutical delivery agents in the treatment of disease.
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