scholarly journals Guiding protein delivery into live cells using DNA-programmed membrane fusion

2018 ◽  
Vol 9 (27) ◽  
pp. 5967-5975 ◽  
Author(s):  
Lele Sun ◽  
Yanjing Gao ◽  
Yaoguang Wang ◽  
Qin Wei ◽  
Jiye Shi ◽  
...  
Keyword(s):  
Membrane Fusion ◽  
Protein Delivery ◽  
Intracellular Delivery ◽  
Live Cells ◽  
Fusion Strategy

A DNA-programmed membrane fusion strategy for directing intracellular delivery of proteins into live cells.

scholarly journals A boronic acid–rich dendrimer with robust and unprecedented efficiency for cytosolic protein delivery and CRISPR-Cas9 gene editing

2019 ◽  
Vol 5 (6) ◽  
pp. eaaw8922 ◽  
Author(s):  
Chongyi Liu ◽  
Tao Wan ◽  
Hui Wang ◽  
Song Zhang ◽  
Yuan Ping ◽  
...  
Keyword(s):  
Boronic Acid ◽  
Protein Delivery ◽  
High Efficiency ◽  
Intracellular Delivery ◽  
Cytosolic Protein ◽  
Native Proteins ◽  
Cytosolic Delivery ◽  
Cas9 Protein ◽  
Cargo Proteins ◽  
Positively Charged

Cytosolic protein delivery is of central importance for the development of protein-based biotechnologies and therapeutics; however, efficient intracellular delivery of native proteins remains a challenge. Here, we reported a boronic acid–rich dendrimer with unprecedented efficiency for cytosolic delivery of native proteins. The dendrimer could bind with both negatively and positively charged proteins and efficiently delivered 13 cargo proteins into the cytosol of living cells. All the delivered proteins kept their bioactivities after cytosolic delivery. The dendrimer ensures efficient intracellular delivery of Cas9 protein into various cell lines and showed high efficiency in CRISPR-Cas9 genome editing. The rationally designed boronic acid–rich dendrimer permits the development of an efficient platform with high generality for the delivery of native proteins.

A versatile pH-responsive platform for intracellular protein delivery using calcium phosphate nanoparticles

2015 ◽  
Vol 3 (47) ◽  
pp. 9115-9121 ◽  
Author(s):  
Bingru Zeng ◽  
Hongdong Shi ◽  
Yangzhong Liu
Keyword(s):  
Calcium Phosphate ◽  
Protein Delivery ◽  
Intracellular Delivery ◽  
Endosomal Escape ◽  
Ph Responsive ◽  
Intracellular Protein ◽  
Calcium Phosphate Nanoparticles ◽  
Intracellular Protein Delivery

A highly biocompatible nanoplatform for the intracellular delivery of different proteins, exhibiting pH-responsive release and efficient endosomal escape.

Intracellular Delivery of Nanobodies for Imaging of Target Proteins in Live Cells

2016 ◽  
Vol 34 (1) ◽  
pp. 161-174 ◽  
Author(s):  
Ruth Röder ◽  
Jonas Helma ◽  
Tobias Preiß ◽  
Joachim O. Rädler ◽  
Heinrich Leonhardt ◽  
...  
Keyword(s):  
Intracellular Delivery ◽  
Live Cells ◽  
Target Proteins

scholarly journals Detecting and controlling dye effects in single-virus fusion experiments

2019 ◽  
Author(s):  
R. J. Rawle ◽  
A. M. Villamil Giraldo ◽  
S. G. Boxer ◽  
P. M. Kasson
Keyword(s):  
Membrane Fusion ◽  
Fluorescent Dyes ◽  
Light Exposure ◽  
Viral Proteins ◽  
Live Cells ◽  
Dependent Manner ◽  
Viral Fusion ◽  
Texas Red ◽  
Kinetics Of ◽  
Cellular Fusion

AbstractFluorescent dye-dequenching assays provide a powerful and versatile means to monitor membrane fusion events. They have been used in bulk assays, for measuring single events in live cells, and for detailed analysis of fusion kinetics for liposomal, viral, and cellular fusion processes; however, the dyes used also have the potential to perturb membrane fusion. Here, using single-virus measurements of influenza membrane fusion, we show that fluorescent membrane probes can alter both the efficiency and the kinetics of lipid mixing in a dye- and illumination-dependent manner. R18, a dye that is commonly used to monitor lipid mixing between membranes, is particularly prone to these effects, while Texas Red is somewhat less sensitive. R18 further undergoes photoconjugation to viral proteins in an illumination-dependent manner that correlates with its inactivation of viral fusion. These results demonstrate how fluorescent probes can perturb measurements of biological activity and provide both data and a method for determining minimally perturbative measurement conditions.Statement of SignificanceFluorescent dyes are powerful tools for labeling membranes and tracking subcellular objects, and fluorescence dequenching has further been used as a sensitive assay for membrane fusion. Here we show how incorporation of membrane dyes can perturb membrane fusion by influenza virus in a light-dependent manner. We provide a strategy to mitigate this by minimizing dye and light exposure. Finally, we show how in some cases these effects can be due to covalent reaction of some dyes with viral proteins upon illumination. These phenomena may be general and should be carefully controlled for in experiments using such labels.

scholarly journals A Chemical-genetics and Nanoparticle Enabled Approach for in vivo Protein Kinase Analysis

2020 ◽  
Author(s):  
Fengqian Chen ◽  
Qi Liu ◽  
Terrell Hilliard ◽  
Tingzeng Wang ◽  
Hongjun Liang ◽  
...  
Keyword(s):  
Protein Kinase ◽  
Protein Kinases ◽  
Intracellular Delivery ◽  
Binding Pocket ◽  
Live Cells ◽  
Genetic Method ◽  
Chemical Genetic ◽  
Methionine Residue ◽  
Two Factors

AbstractThe human kinome contains >500 protein kinases, and regulates up to 30% of the proteome. Kinase study is currently hindered by a lack of in vivo analysis approaches due to two factors: our inability to distinguish the kinase reaction of interest from those of other kinases in live cells and the cell impermeability of the ATP analogs. Herein, we tackled this issue by combining the widely used chemical genetic method developed by Dr. Kevan Shokat and colleagues with nanoparticle-mediated intracellular delivery of the ATP analog. The critical AKT1 protein kinase, which has been successfully studied with the method, was used as our initial prototype. Briefly, enlargement of the ATP binding pocket, by mutating the gate-keeper Methionine residue to a Glycine, prompted the mutant AKT1 to preferentially use the bulky ATP analog N6-Benzyl-ATP-γ-S (A*TPγS) and, thus, differentiating AKT1-catalyzed and other phosphorylation events. The lipid/calcium/phosphate (LCP) nanoparticle was used for efficient intracellular delivery of A*TPγS, overcoming the cell impermeability issue. The mutant, but not wild-type, AKT1 used the delivered A*TPγS for autophosphorylation and phosphorylating its substrates in live cells. Thus, an in vivo protein kinase analysis method has been developed. The strategy should be widely applicable to other protein kinases.

scholarly journals Cationic lipid-mediated intracellular delivery of antibodies into live cells

BioTechniques ◽  
2008 ◽  
Vol 44 (7S) ◽  
pp. vii-xi ◽  
Author(s):  
Claire O. Weill ◽  
Stéphanie Biri ◽  
Patrick Erbacher
Keyword(s):  
Cationic Lipid ◽  
Intracellular Delivery ◽  
Live Cells

scholarly journals Protein delivery into cells using inorganic nanoparticle–protein supramolecular assemblies

2018 ◽  
Vol 47 (10) ◽  
pp. 3421-3432 ◽  
Author(s):  
Federica Scaletti ◽  
Joseph Hardie ◽  
Yi-Wei Lee ◽  
David C. Luther ◽  
Moumita Ray ◽  
...  
Keyword(s):  
Protein Delivery ◽  
Intracellular Delivery ◽  
Supramolecular Assemblies ◽  
Inorganic Nanoparticle

Direct intracellular delivery of proteins using inorganic nanoparticle–protein supramolecular assemblies.

A new technique for reversible permeabilization of live cells for intracellular delivery of quantum dots

2013 ◽  
Vol 24 (20) ◽  
pp. 205101 ◽  
Author(s):  
Krishnakiran Medepalli ◽  
Bruce W Alphenaar ◽  
Robert S Keynton ◽  
Palaniappan Sethu
Keyword(s):  
Quantum Dots ◽  
Intracellular Delivery ◽  
New Technique ◽  
Live Cells ◽  
Reversible Permeabilization ◽  
A New Technique

scholarly journals Protein delivery into live cells by incubation with an endosomolytic agent

2014 ◽  
Vol 11 (8) ◽  
pp. 861-867 ◽  
Author(s):  
Alfredo Erazo-Oliveras ◽  
Kristina Najjar ◽  
Laila Dayani ◽  
Ting-Yi Wang ◽  
Gregory A Johnson ◽  
...  
Keyword(s):  
Protein Delivery ◽  
Live Cells ◽  
Endosomolytic Agent
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