scholarly journals Protein Delivery: Intracellular Delivery of His‐Tagged Genome‐Editing Proteins Enabled by Nitrilotriacetic Acid–Containing Lipidoid Nanoparticles (Adv. Healthcare Mater. 6/2019)

2019 ◽  
Vol 8 (6) ◽  
pp. 1970021
Author(s):  
Yamin Li ◽  
Alice Chukun Li ◽  
Qiaobing Xu
Keyword(s):  
Genome Editing ◽  
Protein Delivery ◽  
Intracellular Delivery ◽  
Nitrilotriacetic Acid

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.

Combinatorial library of chalcogen-containing lipidoids for intracellular delivery of genome-editing proteins

Biomaterials ◽  
2018 ◽  
Vol 178 ◽  
pp. 652-662 ◽  
Author(s):  
Yamin Li ◽  
Tao Yang ◽  
Yingjie Yu ◽  
Nicola Shi ◽  
Liu Yang ◽  
...  
Keyword(s):  
Genome Editing ◽  
Combinatorial Library ◽  
Intracellular Delivery

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.

scholarly journals Efficient delivery of genome-editing proteins using bioreducible lipid nanoparticles

2016 ◽  
Vol 113 (11) ◽  
pp. 2868-2873 ◽  
Author(s):  
Ming Wang ◽  
John A. Zuris ◽  
Fantao Meng ◽  
Holly Rees ◽  
Shuo Sun ◽  
...  
Keyword(s):  
Genome Editing ◽  
Protein Delivery ◽  
Lipid Nanoparticles ◽  
Gene Recombination ◽  
Functional Protein ◽  
Electrostatic Assembly ◽  
Efficient Delivery ◽  
Cultured Human Cells ◽  
Rna Complexes

A central challenge to the development of protein-based therapeutics is the inefficiency of delivery of protein cargo across the mammalian cell membrane, including escape from endosomes. Here we report that combining bioreducible lipid nanoparticles with negatively supercharged Cre recombinase or anionic Cas9:single-guide (sg)RNA complexes drives the electrostatic assembly of nanoparticles that mediate potent protein delivery and genome editing. These bioreducible lipids efficiently deliver protein cargo into cells, facilitate the escape of protein from endosomes in response to the reductive intracellular environment, and direct protein to its intracellular target sites. The delivery of supercharged Cre protein and Cas9:sgRNA complexed with bioreducible lipids into cultured human cells enables gene recombination and genome editing with efficiencies greater than 70%. In addition, we demonstrate that these lipids are effective for functional protein delivery into mouse brain for gene recombination in vivo. Therefore, the integration of this bioreducible lipid platform with protein engineering has the potential to advance the therapeutic relevance of protein-based genome editing.

Nanoscale Metal-organic Frameworks for Intracellular Delivery of CRISPR/Cas9 Genome Editing Machinery

2021 ◽  
Author(s):  
Qizhen Zheng ◽  
Wenting Li ◽  
Lanqun Mao ◽  
Ming Wang
Keyword(s):  
Genome Editing ◽  
Dna Sequence ◽  
High Efficiency ◽  
Genetic Disorders ◽  
Metal Organic Frameworks ◽  
Intracellular Delivery ◽  
Metal Organic

The discovery of CRISPR/Cas9 genome-editing technology enables the precise manipulation of DNA sequence of mammalians for treating genetic disorders. Despite its high efficiency for genome editing, the introduction of CRISPR/Cas9...

scholarly journals Integrating Combinatorial Lipid Nanoparticle and Chemically Modified Protein for Intracellular Delivery and Genome Editing

2018 ◽  
Vol 52 (3) ◽  
pp. 665-675 ◽  
Author(s):  
Jin Chang ◽  
Xianghan Chen ◽  
Zachary Glass ◽  
Feng Gao ◽  
Lanqun Mao ◽  
...  
Keyword(s):  
Genome Editing ◽  
Intracellular Delivery ◽  
Chemically Modified ◽  
Lipid Nanoparticle

scholarly journals Protocol for assessment of the efficiency of CRISPR/Cas RNP delivery to different types of target cells

PLoS ONE ◽  
2021 ◽  
Vol 16 (11) ◽  
pp. e0259812
Author(s):  
Marina A. Tyumentseva ◽  
Aleksandr I. Tyumentsev ◽  
Vasiliy G. Akimkin
Keyword(s):  
Genome Editing ◽  
Protein Delivery ◽  
Imaging Techniques ◽  
Target Cells ◽  
Primary Cells ◽  
Expression And Purification ◽  
Delivery Methods ◽  
Protein Expression And Purification ◽  
Different Types ◽  
Reversible Permeabilization

Background Delivery of CRISPR/Cas RNPs to target cells still remains the biggest bottleneck to genome editing. Many efforts are made to develop efficient CRISPR/Cas RNP delivery methods that will not affect viability of target cell dramatically. Popular current methods and protocols of CRISPR/Cas RNP delivery include lipofection and electroporation, transduction by osmocytosis and reversible permeabilization and erythrocyte-based methods. Methods In this study we will assess the efficiency and optimize current CRISPR/Cas RNP delivery protocols to target cells. We will conduct our work using molecular cloning, protein expression and purification, cell culture, flow cytometry (immunocytochemistry) and cellular imaging techniques. Discussion This will be the first extensive comparative study of popular current methods and protocols of CRISPR/Cas RNP delivery to human cell lines and primary cells. All protocols will be optimized and characterized using the following criteria i) protein delivery and genome editing efficacy; ii) viability of target cells after delivery (post-transduction recovery); iii) scalability of delivery process; iv) cost-effectiveness of the delivery process and v) intellectual property rights. Some methods will be considered ‘research-use only’, others will be recommended for scaling and application in the development of cell-based therapies.

An intracellular protein delivery platform based on glutathione-responsive protein nanocapsules

2016 ◽  
Vol 52 (93) ◽  
pp. 13608-13611 ◽  
Author(s):  
Jie Li ◽  
Linlin Zhang ◽  
Yang Liu ◽  
Jing Wen ◽  
Di Wu ◽  
...  
Keyword(s):  
Protein Delivery ◽  
Intracellular Delivery ◽  
Intracellular Protein ◽  
Disulfide Crosslinking ◽  
Efficient Strategy ◽  
Protein Polymer ◽  
Delivery Platform ◽  
The Stability ◽  
Intracellular Protein Delivery ◽  
Polymer Assembly

We reported an efficient strategy for the intracellular delivery of proteins based on assembling proteins with a self-crosslinkable polymer. The disulfide-crosslinking structure enhances the stability of the protein–polymer assembly, and also allows effective dissociation of the assembly in response to glutathione.

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