GFP-complementation Assay to Detect Functional CPP and Protein Delivery into Living Cells

Overview

Journal Sci Rep

Specialty Science

Date 2015 Dec 17

PMID 26671759

Citations 30

Authors

Nadia Milech

Brooke A C Longville

Paula T Cunningham

Marie N Scobie

Heique M 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

Richard M Hopkins

Affiliations

Soon will be listed here.

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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