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

2015 ◽  
Vol 5 (1) ◽  
Author(s):  
Nadia Milech ◽  
Brooke AC Longville ◽  
Paula T Cunningham ◽  
Marie N Scobie ◽  
Heique M Bogdawa ◽  
...  
Keyword(s):  
Protein Delivery ◽  
Living Cells ◽  
Complementation Assay

scholarly journals Nanopore-mediated protein delivery enabling three-color single-molecule tracking in living cells

2021 ◽  
Vol 118 (5) ◽  
pp. e2012229118
Author(s):  
Zhongwen Chen ◽  
Yuhong Cao ◽  
Chun-Wei Lin ◽  
Steven Alvarez ◽  
Dongmyung Oh ◽  
...  
Keyword(s):  
Single Molecule ◽  
Protein Delivery ◽  
Chemical Properties ◽  
Practical Method ◽  
Living Cells ◽  
Egfr Signaling ◽  
Single Molecule Tracking ◽  
Ras Activation ◽  
And Chemical Properties

Multicolor single-molecule tracking (SMT) provides a powerful tool to mechanistically probe molecular interactions in living cells. However, because of the limitations in the optical and chemical properties of currently available fluorophores and the multiprotein labeling strategies, intracellular multicolor SMT remains challenging for general research studies. Here, we introduce a practical method employing a nanopore-electroporation (NanoEP) technique to deliver multiple organic dye-labeled proteins into living cells for imaging. It can be easily expanded to three channels in commercial microscopes or be combined with other in situ labeling methods. Utilizing NanoEP, we demonstrate three-color SMT for both cytosolic and membrane proteins. Specifically, we simultaneously monitored single-molecule events downstream of EGFR signaling pathways in living cells. The results provide detailed resolution of the spatial localization and dynamics of Grb2 and SOS recruitment to activated EGFR along with the resultant Ras activation.

scholarly journals Rapid protein delivery to living cells for biomolecular investigation

2021 ◽  
Vol 570 ◽  
pp. 82-88
Author(s):  
Francesco Torricella ◽  
Alessio Bonucci ◽  
Panagis Polykretis ◽  
Francesca Cencetti ◽  
Lucia Banci
Keyword(s):  
Protein Delivery ◽  
Living Cells

scholarly journals Efficient Protein Transduction Method Using Cationic Peptides and Lipids

2011 ◽  
Vol 2011 ◽  
pp. 1-6 ◽  
Author(s):  
Kei Yamaguchi ◽  
Mari Inoue ◽  
Naoki Goshima
Keyword(s):  
Protein Delivery ◽  
Cationic Lipid ◽  
Living Cells ◽  
Protein Transduction ◽  
Cationic Peptides ◽  
Proton Sponge ◽  
Protein Transduction Domains ◽  
Cargo Proteins ◽  
Endosomal Vesicles ◽  
Novel Protein

Cationic peptides termed protein transduction domains (PTDs) have been shown to cross biological membranes efficiently. However, proteins transduced by PTDs become entrapped within the endosomal vesicles and are not delivered into organelles. We have developed a novel protein delivery system to enhance the proton sponge effect, which results in rupture of the endosomes, by using a mixture of Wr-T transporter peptide and a commercially available cationic lipid reagent. This peptide and cationic lipid reagent mixture efficiently delivers a variety of cargo proteins into living cells by releasing them from the endosomes.

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