Jove
Visualize
Contact Us
JoVE
x logofacebook logolinkedin logoyoutube logo
ABOUT JoVE
OverviewLeadershipBlogJoVE Help Center
AUTHORS
Publishing ProcessEditorial BoardScope & PoliciesPeer ReviewFAQSubmit
LIBRARIANS
TestimonialsSubscriptionsAccessResourcesLibrary Advisory BoardFAQ
RESEARCH
JoVE JournalMethods CollectionsJoVE Encyclopedia of ExperimentsArchive
EDUCATION
JoVE CoreJoVE BusinessJoVE Science EducationJoVE Lab ManualFaculty Resource CenterFaculty Site
Terms & Conditions of Use
Privacy Policy
Policies

Related Concept Videos

Reporter Genes02:11

Reporter Genes

13.8K
Reporter genes are a type of protein-coding gene that are often tagged to a gene of interest. Once inside a target cell, reporter genes usually produce visually identifiable characteristics like fluorescence and luminescence when expressed along with the gene of interest. Thus, reporter genes “report” the presence or absence of genes of interest in an organism, determine the gene expression pattern, or track the physical location of a DNA segment or protein in the cell.
13.8K
Protein Dynamics in Living Cells01:19

Protein Dynamics in Living Cells

2.8K
Different fluorescence-based techniques are used to study the protein dynamics in living cells. These techniques include FRAP, FRET, and PET.
Fluorescent recovery after photobleaching (FRAP) is a fluorescent-protein-based detection technique used to quantify protein movement rates within the cell. This method exposes a small portion of the cell to an intense laser beam. The laser beam causes permanent photobleaching of the fluorophore-tagged proteins in the exposed region. As the bleached...
2.8K

You might also read

Related Articles

Articles linked to this work by shared authors, journal, and citation graph.

Sort by
Same author

RETRACTED: Wang et al. HSP70-eIF4G Interaction Promotes Protein Synthesis and Cell Proliferation in Hepatocellular Carcinoma. <i>Cancers</i> 2020, <i>12</i>, 3410.

Cancers·2026
Same author

Longitudinal surveillance of group A streptococcal pharyngitis and impetigo in remote Western Australian school children informs acute rheumatic fever prevention.

PLOS global public health·2025
Same author

Diclofenac and acetaminophen dim the acute-phase response but amplify expression of the iron regulator hepcidin in liver cancer cells.

Cell systems·2025
Same author

Physical and motivational effects of Exergames in healthy adults-A scoping review.

PloS one·2025
Same author

Reappraisal of liver resection as an alternative to transplantation in locally advanced hepatoblastoma: A systematic review and analysis of pooled individual patient data.

Pediatric blood & cancer·2024
Same author

A20 haploinsufficiency disturbs immune homeostasis and drives the transformation of lymphocytes with permissive antigen receptors.

Science advances·2024

Related Experiment Video

Updated: Mar 28, 2026

Optimizing the Genetic Incorporation of Chemical Probes into GPCRs for Photo-crosslinking Mapping and Bioorthogonal Chemistry in Live Mammalian Cells
14:02

Optimizing the Genetic Incorporation of Chemical Probes into GPCRs for Photo-crosslinking Mapping and Bioorthogonal Chemistry in Live Mammalian Cells

Published on: April 9, 2018

9.2K

GFP-complementation assay to detect functional CPP and protein delivery into living cells.

Nadia Milech1, Brooke A C Longville1, Paula T Cunningham1

  • 1Telethon Kids Institute &Centre for Child Health Research, The University of Western Australia, Drug Discovery Group. West Perth, 6872, Australia.

Scientific Reports
|December 17, 2015
PubMed
Summary

A new assay called Split-complementation Endosomal Escape (SEE) directly visualizes cell-penetrating peptide (CPP) delivery into the cytoplasm. This method overcomes limitations of current assays, enabling better characterization of CPPs for drug delivery.

More Related Videos

Assay to Measure Nucleocytoplasmic Transport in Real Time within Motor Neuron-like NSC-34 Cells
08:53

Assay to Measure Nucleocytoplasmic Transport in Real Time within Motor Neuron-like NSC-34 Cells

Published on: May 16, 2017

9.3K
Effect of Fluorescent Proteins on Fusion Partners Using Polyglutamine Toxicity Assays in Yeast
09:23

Effect of Fluorescent Proteins on Fusion Partners Using Polyglutamine Toxicity Assays in Yeast

Published on: November 28, 2018

7.4K

Related Experiment Videos

Last Updated: Mar 28, 2026

Optimizing the Genetic Incorporation of Chemical Probes into GPCRs for Photo-crosslinking Mapping and Bioorthogonal Chemistry in Live Mammalian Cells
14:02

Optimizing the Genetic Incorporation of Chemical Probes into GPCRs for Photo-crosslinking Mapping and Bioorthogonal Chemistry in Live Mammalian Cells

Published on: April 9, 2018

9.2K
Assay to Measure Nucleocytoplasmic Transport in Real Time within Motor Neuron-like NSC-34 Cells
08:53

Assay to Measure Nucleocytoplasmic Transport in Real Time within Motor Neuron-like NSC-34 Cells

Published on: May 16, 2017

9.3K
Effect of Fluorescent Proteins on Fusion Partners Using Polyglutamine Toxicity Assays in Yeast
09:23

Effect of Fluorescent Proteins on Fusion Partners Using Polyglutamine Toxicity Assays in Yeast

Published on: November 28, 2018

7.4K

Area of Science:

  • Biochemistry
  • Cell Biology
  • Drug Delivery

Background:

  • Cell-penetrating peptides (CPPs) are crucial for delivering diverse cargoes into cells.
  • Current CPP uptake assays have limitations, including indirect read-outs and difficulty distinguishing true cytoplasmic delivery from endosomal trapping.

Purpose of the Study:

  • To develop a direct assay for visualizing and quantifying true cytoplasmic delivery of protein cargoes mediated by CPPs.
  • To overcome the limitations of existing CPP activity assays.

Main Methods:

  • Development of the Split-complementation Endosomal Escape (SEE) assay, a split-GFP-based platform.
  • Utilizing SEE to directly visualize protein translocation into the cytoplasm at biologically relevant concentrations.

Main Results:

  • The SEE assay provides a direct visualization of cytoplasmic delivery, minimizing background noise.
  • The assay is adaptable to various cell lines (transient and stable) and amenable to high-throughput screening.
  • SEE effectively discriminates true internalization from endosomal sequestration.

Conclusions:

  • The SEE assay is the first direct method to visualize true cytoplasmic delivery of proteins by CPPs.
  • This platform offers a robust tool for studying CPP transduction mechanisms, cellular imaging, and characterizing novel CPPs for pharmaceutical applications.