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Transduction Methods For Cytosolic Delivery Of Proteins And Bioconjugates Into Living Cells
M. Chiper, K. Niederreither, G. Zuber
Published 2018 · Chemistry, Medicine
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The human organism and its constituting cells rely on interplay between multiple proteins exerting specific functions. Progress in molecular biotechnologies has facilitated the production of recombinant proteins. When administrated to patients, recombinant proteins can provide important healthcare benefits. To date, most therapeutic proteins must act from the extracellular environment, with their targets being secreted modulators or extracellular receptors. This is because proteins cannot passively diffuse across the plasma membrane into the cytosol. To expand the scope of action of proteins for cytosolic targets (representing more than 40% of the genome) effective methods assisting protein cytosolic entry are being developed. To date, direct protein delivery is extremely tedious and inefficient in cultured cells, even more so in animal models of pathology. Novel techniques are changing this limitation, as recently developed in vitro methods can robustly convey large amount of proteins into cell cultures. Moreover, advances in protein formulation or protein conjugates are slowly, but surely demonstrating efficiency for targeted cytosolic entry of functional protein in vivo in tumor xenograft models. In this review, various methods and recently developed techniques for protein transport into cells are summarized. They are put into perspective to address the challenges encountered during delivery.
This paper references
Electroporation of antibodies, DNA, and other macromolecules into cells: a highly efficient method.
S. Baron (2000)
Electroporation-based technologies for medicine: principles, applications, and challenges.
M. Yarmush (2014)
Comparative analysis of the methods of drug and protein delivery for the treatment of cancer, genetic diseases and diagnostics
R. Todorova (2011)
Highly Efficient Delivery of Functional Cargoes by a Novel Cell-Penetrating Peptide Derived from SP140-Like Protein.
H. Wang (2016)
Nucleic acid-mediated intracellular protein delivery by lipid-like nanoparticles.
Ahmed A. Eltoukhy (2014)
Mechanisms of femtosecond laser nanosurgery of cells and tissues
A. Vogel (2005)
Phosphorylation by p34cdc2 regulates spindle association of human Eg5, a kinesin-related motor essential for bipolar spindle formation in vivo
Anne Slangy (1995)
Reduction of Infarct Size by the Therapeutic Protein TAT-Ndi1 In Vivo
R. Mentzer (2014)
Reprogramming of somatic cells via TAT-mediated protein transduction of recombinant factors.
Hui Bin Zhang (2012)
Cellular manipulation of human embryonic stem cells by TAT-PDX1 protein transduction.
Y. Kwon (2005)
Intracellular protein delivery with a dimerizable amphiphile for improved complex stability and prolonged protein release in the cytoplasm of adherent cell lines.
Deniz Dalkara (2006)
Current Understanding of Direct Translocation of Arginine-Rich Cell-Penetrating Peptides and Its Internalization Mechanisms.
T. Takeuchi (2016)
Protein Delivery System Containing a Nickel-Immobilized Polymer for Multimerization of Affinity-Purified His-Tagged Proteins Enhances Cytosolic Transfer.
V. Postupalenko (2015)
Cationic lipid-mediated delivery of proteins enables efficient protein-based genome editing in vitro and in vivo
J. Zuris (2015)
Massively parallel delivery of large cargo into mammalian cells with light pulses
Yi-Chien Wu (2015)
Tat-mediated delivery of heterologous proteins into cells.
S. Fawell (1994)
Peptide-Mediated Cellular Delivery of Oligonucleotide-Based Therapeutics In Vitro: Quantitative Evaluation of Overall Efficacy Employing Easy to Handle Reporter Systems
S.D Laufer (2008)
A general strategy for generating intact, full-length IgG antibodies that penetrate into the cytosol of living cells
D. Choi (2014)
Endosome disruption enhances the functional nuclear delivery of Tat-fusion proteins.
N. Caron (2004)
Degradable polymeric nanocapsule for efficient intracellular delivery of a high molecular weight tumor-selective protein complex
Muxun Zhao (2013)
Promises and Pitfalls of Intracellular Delivery of Proteins
Ailing Fu (2014)
Exosomes as therapeutic drug carriers and delivery vehicles across biological membranes: current perspectives and future challenges
Dinh Ha (2016)
Monomolecular DNA nanoparticles for intravenous delivery of genes.
Chandrashekhar Chittimalla (2005)
Live cell immunogold labelling of RNA polymerase II
Igor Orlov (2015)
Intracytoplasmic delivery of anionic proteins.
Deniz Dalkara (2004)
Intracellular delivery of the PTEN protein using cationic lipidoids for cancer therapy.
Sarah Altinoglu (2016)
Alveolar wall apoptosis causes lung destruction and emphysematous changes.
K. Aoshiba (2003)
Intracellular Delivery of Nanobodies for Imaging of Target Proteins in Live Cells
Ruth Röder (2016)
Sequence segregation improves non‐covalent protein delivery
Federica Sgolastra (2017)
Human‐derived fusogenic peptides for the intracellular delivery of proteins
Kei Sudo (2017)
Gramicidin A-based peptide vector for intracellular protein delivery.
Tatiana B Stoilova (2008)
Early Endosomal Escape of a Cyclic Cell-Penetrating Peptide Allows Effective Cytosolic Cargo Delivery
Ziqing Qian (2014)
Serum insensitive, intranuclear protein delivery by the multipurpose cationic lipid SAINT-2.
B. V. D. van der Gun (2007)
Enhancing Endosomal Escape for Intracellular Delivery of Macromolecular Biologic Therapeutics
P. Lönn (2016)
Delivery of antibodies to the cytosol
Andrea L. J. Marschall (2014)
Tailoring nanocarriers for intracellular protein delivery.
Z. Gu (2011)
Cloning and Characterization of the ζ-Carotene Desaturase Gene from Chlorella protothecoides CS-41
M. Li (2011)
Intracellular Delivery of Proteins with a New Lipid-mediated Delivery System*
O. Zelphati (2001)
Global quantification of mammalian gene expression control
Björn Schwanhäusser (2011)
Efficient Protein Transduction Method Using Cationic Peptides and Lipids
K. Yamaguchi (2011)
The biodistribution of gold nanoparticles designed for renal clearance.
Christophe Alric (2013)
Delivery of proteins into living cells by reversible membrane permeabilization with streptolysin-O
I. Walev (2001)
A human single chain transbody specific to matrix protein (M1) interferes with the replication of influenza A virus.
Ornnuthchar Poungpair (2010)
Camelid single-domain antibodies: A versatile tool for in vivo imaging of extracellular and intracellular brain targets.
Tengfei Li (2016)
Protein nanocapsule weaved with enzymatically degradable polymeric network.
Z. Gu (2009)
Efficient delivery of bioactive antibodies into the cytoplasm of living cells by charge-conversional polyion complex micelles.
Yan Lee (2010)
Covalent attachment of cyclic TAT peptides to GFP results in protein delivery into live cells with immediate bioavailability.
N. Nischan (2015)
Nanomotor-Enabled pH-Responsive Intracellular Delivery of Caspase-3: Toward Rapid Cell Apoptosis.
B. Esteban-Fernández de Ávila (2017)
Targeted release of transcription factors for human cell reprogramming by ZEBRA cell-penetrating peptide.
Benjamin Caulier (2017)
N. Nischan (1950)
In Vivo protein transduction: Delivery of PEP-1-SOD1 fusion protein into myocardium efficiently protects against ischemic insult
Y. Zhang (2009)
Quantitative measurement of delivery and gene silencing activities of siRNA polyplexes containing pyridylthiourea-grafted polyethylenimines.
S. Pinel (2014)
Senescence-associated (beta)-galactosidase reflects an increase in lysosomal mass during replicative ageing of human endothelial cells.
D. J. Kurz (2000)
Rational Design of a Polymer with Robust Efficacy for Intracellular Protein and Peptide Delivery.
H. Chang (2017)
In Vivo Delivery of a Bcl-xL Fusion Protein Containing the TAT Protein Transduction Domain Protects against Ischemic Brain Injury and Neuronal Apoptosis
G. Cao (2002)
Cytoarchitecture and physical properties of cytoplasm: volume, viscosity, diffusion, intracellular surface area.
K. Luby-Phelps (2000)
Protein delivery of caspase-3 induces cell death in malignant C6 glioma, primary astrocytes and immortalized and primary brain capillary endothelial cells
B. Zassler (2004)
A versatile vector for gene and oligonucleotide transfer into cells in culture and in vivo: polyethylenimine.
O. Boussif (1995)
Transducible TAT-HA fusogenic peptide enhances escape of TAT-fusion proteins after lipid raft macropinocytosis
J. Wadia (2004)
Lipo-Oligomer Nanoformulations for Targeted Intracellular Protein Delivery.
P. Zhang (2017)
Enhanced Intracellular Protein Transduction by Sequence Defined Tetra‐Oleoyl Oligoaminoamides Targeted for Cancer Therapy
P. Zhang (2015)
Cell-penetrating TAT peptide in drug delivery systems: Proteolytic stability requirements
E. Koren (2011)
Magnetically Guided Protein Transduction by Hybrid Nanogel Chaperones with Iron Oxide Nanoparticles.
R. Kawasaki (2016)
EGFR and CD44 Dual-Targeted Multifunctional Hyaluronic Acid Nanogels Boost Protein Delivery to Ovarian and Breast Cancers In Vitro and In Vivo.
J. Chen (2017)
Cytosolic antibody delivery by lipid-sensitive endosomolytic peptide.
Misao Akishiba (2017)
Potent synergy of dual antitumor peptides for growth suppression of human glioblastoma cell lines
E. Kondo (2008)
Antibodies inside of a cell can change its outside: Can intrabodies provide a new therapeutic paradigm?
Andrea L. J. Marschall (2016)
Self‐Assembling Polyethylenimine Derivatives Mediate Efficient siRNA Delivery in Mammalian Cells
Gaelle Creusat (2008)
Active Targeting of the Nucleus Using Nonpeptidic Boronate Tags.
Rui Tang (2017)
A peptide-based carrier for intracellular delivery of proteins into malignant glial cells in vitro.
C. Wu (2008)
Protein import into the cell nucleus.
C. Dingwall (1986)
Exosomes: current knowledge of their composition, biological functions, and diagnostic and therapeutic potentials.
A. Vlassov (2012)
pH-Reversible Cationic RNase A Conjugates for Enhanced Cellular Delivery and Tumor Cell Killing.
Xiaowen Liu (2016)
Internalization via Antennapedia protein transduction domain of an scFv antibody toward c‐Myc protein
C. Avignolo (2008)
Autophagy proteins regulate innate immune responses by inhibiting the release of mitochondrial DNA mediated by the NALP3 inflammasome.
K. Nakahira (2011)
Tissue-based map of the human proteome
M. Uhlén (2015)
Cell-Penetrating Peptide-Mediated Delivery of TALEN Proteins via Bioconjugation for Genome Engineering
Jia Liu (2014)
Gold nanoparticle-DNA aptamer composites as a universal carrier for in vivo delivery of biologically functional proteins.
Sangmi Ryou (2014)
Quantitative tracking of protein trafficking to the nucleus using cytosolic protein delivery by nanoparticle-stabilized nanocapsules.
M. Ray (2015)
The combined transduction of copper, zinc-superoxide dismutase and catalase mediated by cell-penetrating peptide, PEP-1, to protect myocardium from ischemia-reperfusion injury
Guang-Qing Huang (2011)
Intracellular delivery of a membrane-impermeable enzyme in active form using functionalized gold nanoparticles.
P. Ghosh (2010)
Membranotropic Cell Penetrating Peptides: The Outstanding Journey
A. Falanga (2015)
History of Polymeric Gene Delivery Systems
P. Zhang (2017)
Self‐aggregating 1.8 kDa polyethylenimines with dissolution switch at endosomal acidic pH are delivery carriers for plasmid DNA, mRNA, siRNA and exon‐skipping oligonucleotides
M. Chiper (2017)
Stimuli-responsive nanomaterials for therapeutic protein delivery.
Y. Lu (2014)
Targeted Intracellular Delivery of Proteins with Spatial and Temporal Control
Demosthenes P. Morales (2015)
Long-circulating heparin-functionalized magnetic nanoparticles for potential application as a protein drug delivery platform.
J. Zhang (2013)
PEP and CADY-mediated delivery of fluorescent peptides and proteins into living cells.
Laetitia Kurzawa (2010)
Exosome engineering for efficient intracellular delivery of soluble proteins using optically reversible protein–protein interaction module
Nambin Yim (2016)
In vivo protein transduction: delivery of a biologically active protein into the mouse.
S. Schwarze (1999)
Cell penetrating peptides as an innovative approach for drug delivery; then, present and the future
Santosh K Bashyal (2016)
Reduction and pH dual-bioresponsive crosslinked polymersomes for efficient intracellular delivery of proteins and potent induction of cancer cell apoptosis.
H. Sun (2014)
Polymers for siRNA delivery: inspired by viruses to be targeted, dynamic, and precise.
E. Wagner (2012)
Highly efficient delivery of functional cargoes by the synergistic effect of GAG binding motifs and cell-penetrating peptides
James E. Dixon (2016)
Sonodynamic cancer therapy: a non-invasive and repeatable approach using low-intensity ultrasound with a sonosensitizer.
H. Shibaguchi (2011)
Superparamagnetic iron oxide contrast agents: physicochemical characteristics and applications in MR imaging
Y. Wang (2014)
A Comprehensive Model for the Cellular Uptake of Cationic Cell‐penetrating Peptides
Falk Duchardt (2007)
Direct delivery of functional proteins and enzymes to the cytosol using nanoparticle-stabilized nanocapsules.
R. Tang (2013)
Star-like oligo-arginyl-maltotriosyl derivatives as novel cell-penetrating enhancers for the intracellular delivery of colloidal therapeutic systems.
Sara Bersani (2012)
Intracellular delivery of functionally active proteins using self-assembling pyridylthiourea-polyethylenimine.
V. Postupalenko (2014)
Clinical potential of electroporation for gene therapy and DNA vaccine delivery
Laure Lambricht (2016)
Quantitative assessment of cellular uptake and cytosolic access of antibody in living cells by an enhanced split GFP complementation assay.
J. Kim (2015)
The taming of the cell penetrating domain of the HIV Tat: myths and realities.
Ashok Chauhan (2007)
K. Yamaguchi (2011)
Efficient delivery of genome-editing proteins using bioreducible lipid nanoparticles
M. Wang (2016)
Targeting antibodies to the cytoplasm
Andrea L. J. Marschall (2011)
Cytosolic delivery of proteins mediated by aldehyde-displaying silica nanoparticles with pH-responsive characteristics
Xuanjun Wu (2012)
Performance of Pyridylthiourea‐Polyethylenimine Polyplex for siRNA‐Mediated Liver Cancer Therapy in Cell Monolayer, Spheroid, and Tumor Xenograft Models
Jean Baptiste Gossart (2017)
Clickable protein nanocapsules for targeted delivery of recombinant p53 protein.
Muxun Zhao (2014)
Cloning, expression, purification, distribution and kinetics characterization of the bacterial beta-galactosidase fused to the cytoplasmic transduction peptide in vitro and in vivo.
Shi-feng Huang (2009)
Synthesis of silica nanoparticles with controllable surface roughness for therapeutic protein delivery.
Yuting Niu (2015)
A peptide carrier for the delivery of biologically active proteins into mammalian cells
M. Morris (2001)
Single-molecule spectroscopy of protein conformational dynamics in live eukaryotic cells
I. Koenig (2015)
Cell-Penetrating Peptides: From Basic Research to Clinics.
Giulia Guidotti (2017)
Suppression of cervical carcinoma cell growth by intracytoplasmic codelivery of anti-oncoprotein E6 antibody and small interfering RNA
Jérôme Courtête (2007)
Delivery of reprogramming factors into fibroblasts for generation of non-genetic induced pluripotent stem cells using a cationic bolaamphiphile as a non-viral vector.
Majad Khan (2013)
Simultaneous adsorptive removal of methylene blue and copper ions from aqueous solution by ferrocene‐modified cation exchange resin
Q. Wang (2014)
Efficient Mini-Transporter for Cytosolic Protein Delivery.
N. Zhang (2016)
Protein Expression Purif
S. Huang (2009)
Intracellular chromobody delivery by mesoporous silica nanoparticles for antigen targeting and visualization in real time
H. Chiu (2016)
Intracellular delivery of proteins in complexes with oligoarginine-modified liposomes and the effect of oligoarginine length.
M. Furuhata (2006)
The Late Endosome and Its Lipid BMP Act as Gateways for Efficient Cytosolic Access of the Delivery Agent dfTAT and Its Macromolecular Cargos.
Alfredo Erazo-Oliveras (2016)
CORRIGENDUM: Ion concentration polarization-based continuous separation device using electrical repulsion in the depletion region
Hyungkook Jeon (2014)
Intracellular delivery of universal proteins using a lysine headgroup containing cationic liposomes: deciphering the uptake mechanism.
S. Sarker (2014)
Protein delivery into live cells by incubation with an endosomolytic agent
Alfredo Erazo-Oliveras (2014)
A novel intracellular protein delivery platform based on single-protein nanocapsules.
Ming Yan (2010)
A vector-free microfluidic platform for intracellular delivery
A. Sharei (2013)
A review of in-vivo optical properties of human tissues and its impact on PDT.
J. Sandell (2011)
Evaluation of Strategies for the Intracellular Delivery of Proteins
D. Ye (2004)
A novel method of DNA transfection by laser microbeam cell surgery
M. Tsukakoshi (1984)
Cytosolic delivery of large proteins using nanoparticle-stabilized nanocapsules.
Rui Tang (2016)
Site-specific protein labeling by Sfp phosphopantetheinyl transferase
J. Yin (2006)
Efficient and High-Speed Transduction of an Antibody into Living Cells Using a Multifunctional Nanocarrier System to Control Intracellular Trafficking.
Y. Yamada (2015)
Co-delivery of protein and small molecule therapeutics using nanoparticle-stabilized nanocapsules.
C. S. Kim (2015)
Targeting endogenous nuclear antigens by electrotransfer of monoclonal antibodies in living cells
Guillaume Freund (2013)
Intracellular delivery of recombinant proteins via gold nanoparticle–DNA aptamer composites is independent of the protein physicochemical properties and cell type
Ji-Hyun Yeom (2017)
Hyaluronic acid modification of RNase A and its intracellular delivery using lipid-like nanoparticles.
Xiaoying Wang (2017)
Multi-watt near-infrared light therapy as a neuroregenerative treatment for traumatic brain injury
Theodore A. Henderson (2016)
Generation of induced pluripotent stem cells using recombinant proteins.
H. Zhou (2009)
General Strategy for Direct Cytosolic Protein Delivery via Protein-Nanoparticle Co-engineering.
Rubul Mout (2017)
Highly efficient delivery of p16 antitumor peptide into aggressive leukemia/lymphoma cells using a novel transporter system.
E. Kondo (2004)
Reprogramming human fibroblasts using HIV-1 TAT recombinant proteins OCT4, SOX2, KLF4 and c-MYC
C. Pan (2009)
Synthetic Protein Mimics for Functional Protein Delivery.
A. Ö. Tezgel (2017)
Towards synthetic viruses.
G. Zuber (2001)
Intracellular delivery of proteins by nanocarriers.
M. Ray (2017)
Specific in vivo knockdown of protein function by intrabodies
Andrea L. J. Marschall (2015)
Potent Delivery of Functional Proteins into Mammalian Cells in Vitro and in Vivo Using a Supercharged Protein
James J. Cronican (2010)
Transduction of full-length TAT fusion proteins into mammalian cells: TAT-p27Kip1 induces cell migration
H. Nagahara (1998)
Cytoplasmic transduction peptide (CTP): new approach for the delivery of biomolecules into cytoplasm in vitro and in vivo.
D. Kim (2006)
The Perforin Pore Facilitates the Delivery of Cationic Cargos*
S. E. Stewart (2014)
Intracellular versus extracellular granzyme B in immunity and disease: challenging the dogma
W. A. Boivin (2009)
Ultrasonically triggered drug delivery: breaking the barrier.
G. Husseini (2014)
Regulation of proliferating cell nuclear antigen during the cell cycle.
G. Morris (1989)
A practical approach for intracellular protein delivery
C. Weill (2007)
Acid-labile traceless click linker for protein transduction.
Kevin Maier (2012)
Emerging landscape of cell penetrating peptide in reprogramming and gene editing.
H. Liu (2016)
Direct Cytosolic Delivery of CRISPR/Cas9-Ribonucleoprotein for Efficient Gene Editing.
Rubul Mout (2017)
Near-Infrared Light Activation of Proteins Inside Living Cells Enabled by Carbon Nanotube-Mediated Intracellular Delivery.
H. Li (2016)
Hyaluronic acid targeting of CD44 for cancer therapy: from receptor biology to nanomedicine*
George Mattheolabakis (2015)
Efficient Intracellular Delivery of Native Proteins
D. S. D'Astolfo (2015)
Polyethylene glycol-based protein nanocapsules for functional delivery of a differentiation transcription factor.
A. Biswas (2012)
Redox compartmentalization in eukaryotic cells.
Y. Go (2008)
Targeting the replisome with transduced monoclonal antibodies triggers lethal DNA replication stress in cancer cells.
D. Desplancq (2016)
Discovery and characterization of a peptide that enhances endosomal escape of delivered proteins in vitro and in vivo.
Margie Li (2015)
HIV is inactivated after transepithelial migration via adult oral epithelial cells but not fetal epithelial cells.
S. Tugizov (2011)
DNA vaccines, electroporation and their applications in cancer treatment
Si-Hyeong Lee (2015)
Resurfaced cell‐penetrating nanobodies: A potentially general scaffold for intracellularly targeted protein discovery
Virginia J Bruce (2016)
Developments in Intralesional Therapy for Metastatic Melanoma.
S. Sloot (2016)
Multifunctional Click Hyaluronic Acid Nanogels for Targeted Protein Delivery and Effective Cancer Treatment in Vivo
J. Chen (2016)
Protection against ischemic brain injury by protein therapeutics
S. Asoh (2002)
Sequence defined disulfide-linked shuttle for strongly enhanced intracellular protein delivery.
Kevin Maier (2012)
This paper is referenced by
Biocompatible gold nanoclusters: synthetic strategies and biomedical prospects.
G. Zuber (2019)
Transient delivery of A-C/EBP protein perturbs differentiation of 3T3-L1 cells and induces preadipocyte marker genes
Nishtha Sharma (2020)
Protein Delivery into the Cell Cytosol using Non-Viral Nanocarriers
Yi-Wei Lee (2019)
Accessing Intracellular Targets through Nanocarrier-Mediated Cytosolic Protein Delivery.
Ritabrita Goswami (2020)
Strategies in the delivery of Cas9 ribonucleoprotein for CRISPR/Cas9 genome editing
S. Zhang (2020)
Synthesis and biological evaluation of 2.4 nm thiolate-protected gold nanoparticles conjugated to Cetuximab for targeting glioblastoma cancer cells via the EGFR.
Nadja Groysbeck (2019)
Protein Nanotherapeutics as an Emerging Modality for Cancer Therapy
Liang Cheng (2018)
Selection of Functional Intracellular Nanobodies
J. Woods (2019)
Co-delivery of doxorubicin and paclitaxel by reduction/pH dual responsive nanocarriers for osteosarcoma therapy
Y. Li (2020)