Designed filamentous cell penetrating peptides: probing supramolecular structure-dependent membrane activity and transfection efficiency

2015 ◽  
Vol 51 (59) ◽  
pp. 11757-11760 ◽  
Author(s):  
Dawei Xu ◽  
Derek Dustin ◽  
Linhai Jiang ◽  
Damien S. K. Samways ◽  
He Dong
Keyword(s):  
Supramolecular Structure ◽  
Transfection Efficiency ◽  
Supramolecular Assembly ◽  
Cell Penetrating Peptides ◽  
Cationic Peptides ◽  
Single Chain ◽  
Membrane Activity ◽  
Highly Efficient ◽  
Cell Penetrating ◽  
Filamentous Cell

In this work, we will demonstrate the supramolecular assembly of single-chain cationic peptides into stable macromolecular filamentous nanostructures and investigate their supramolecular structure-dependent membrane activity for the development of highly efficient therapeutic carriers.

Synergistic effects of conjugating cell penetrating peptides and thiomers on non-viral transfection efficiency

Biomaterials ◽  
2012 ◽  
Vol 33 (7) ◽  
pp. 2321-2326 ◽  
Author(s):  
Deni Rahmat ◽  
Mohammad I. Khan ◽  
Gul Shahnaz ◽  
Duangkamon Sakloetsakun ◽  
Glen Perera ◽  
...  
Keyword(s):  
Transfection Efficiency ◽  
Synergistic Effects ◽  
Cell Penetrating Peptides ◽  
Cell Penetrating ◽  
Viral Transfection

scholarly journals Influence of the Dabcyl group on the cellular uptake of cationic peptides: short oligoarginines as efficient cell-penetrating peptides

Amino Acids ◽  
2021 ◽  
Author(s):  
Ildikó Szabó ◽  
Françoise Illien ◽  
Levente E. Dókus ◽  
Mo’ath Yousef ◽  
Zsuzsa Baranyai ◽  
...  
Keyword(s):  
Cellular Uptake ◽  
Cell Penetrating Peptides ◽  
Antitumor Drugs ◽  
Short Peptides ◽  
Cationic Peptides ◽  
Wide Range ◽  
Cell Penetrating ◽  
Delivery Vehicles ◽  
Direct Delivery ◽  
Diffuse Distribution

AbstractCell-penetrating peptides (CPPs) are promising delivery vehicles. These short peptides can transport wide range of cargos into cells, although their usage has often limitations. One of them is the endosomatic internalisation and thus the vesicular entrapment. Modifications which increases the direct delivery into the cytosol is highly researched area. Among the oligoarginines the longer ones (n > 6) show efficient internalisation and they are well-known members of CPPs. Herein, we describe the modification of tetra- and hexaarginine with (4–((4–(dimethylamino)phenyl)azo)benzoyl) (Dabcyl) group. This chromophore, which is often used in FRET system increased the internalisation of both peptides, and its effect was more outstanding in case of hexaarginine. The modified hexaarginine may enter into cells more effectively than octaarginine, and showed diffuse distribution besides vesicular transport already at low concentration. The attachment of Dabcyl group not only increases the cellular uptake of the cell-penetrating peptides but it may affect the mechanism of their internalisation. Their conjugates with antitumor drugs were studied on different cells and showed antitumor activity.

scholarly journals Improving Membrane Activity and Cargo Delivery Efficacy of a Cell-Penetrating Peptide by Loading with Carboranes

Pharmaceutics ◽  
2021 ◽  
Vol 13 (12) ◽  
pp. 2075
Author(s):  
Tamara Lützenburg ◽  
Nele Burdina ◽  
Matthias S. Scholz ◽  
Ines Neundorf
Keyword(s):  
Secondary Structure ◽  
Cell Penetrating Peptides ◽  
Nucleic Acid Delivery ◽  
Stable Secondary Structure ◽  
Membrane Activity ◽  
Cargo Delivery ◽  
Peptide Interactions ◽  
Cell Penetrating ◽  
A Cell

Cell-penetrating peptides (CPPs) have emerged as versatile tools to increase the intracellular accumulation of different kinds of cargoes. For an efficient cellular uptake and drug delivery, their organization into a distinct and stable secondary structure at the outer surface of the plasma membrane is a hallmark and supports optimal lipid–peptide interactions. Incorporation of hydrophobic moieties, such as carboranes (CBs), has the potential to increase the lipophilicity of peptides, and thus, to facilitate the formation of secondary structures. Herein, we present synthesis and biophysical as well as biological characterization of carborane-CPP conjugates having incorporated one or more CB clusters. Our results highlight the possibility to modulate the secondary structure of CPPs by the addition of CB’s leading to constructs with altered membrane activity and promising use in terms of nucleic acid delivery.

scholarly journals Discovery and Mechanism of Highly Efficient Cyclic Cell-Penetrating Peptides

Biochemistry ◽  
2016 ◽  
Vol 55 (18) ◽  
pp. 2601-2612 ◽  
Author(s):  
Ziqing Qian ◽  
Agnieszka Martyna ◽  
Ryan L. Hard ◽  
Jiang Wang ◽  
George Appiah-Kubi ◽  
...  
Keyword(s):  
Cell Penetrating Peptides ◽  
Highly Efficient ◽  
Cell Penetrating

scholarly journals Internalisation and Biological Activity of Nucleic Acids Delivering Cell-Penetrating Peptide Nanoparticles Is Controlled by the Biomolecular Corona

2021 ◽  
Vol 14 (7) ◽  
pp. 667
Author(s):  
Annely Lorents ◽  
Maria Maloverjan ◽  
Kärt Padari ◽  
Margus Pooga
Keyword(s):  
Nucleic Acid ◽  
Nucleic Acids ◽  
Transfection Efficiency ◽  
Protein Corona ◽  
Blood Stream ◽  
Cell Penetrating Peptides ◽  
Biological Efficiency ◽  
Cell Penetrating

Nucleic acid molecules can be transferred into cells to alter gene expression and, thus, alleviate certain pathological conditions. Cell-penetrating peptides (CPPs) are vectors that can be used for transfecting nucleic acids as well as many other compounds. CPPs associate nucleic acids non-covalently, forming stable nanoparticles and providing efficient transfection of cells in vitro. However, in vivo, expected efficiency is achieved only in rare cases. One of the reasons for this discrepancy is the formation of protein corona around nanoparticles, once they are exposed to a biological environment, e.g., blood stream. In this study, we compared protein corona of CPP-nucleic acid nanoparticles formed in the presence of bovine, murine and human serum. We used Western blot and mass-spectrometry to identify the major constituents of protein corona forming around nanoparticles, showing that proteins involved in transport, haemostasis and complement system are its major components. We investigated physical features of nanoparticles and measured their biological efficiency in splice-correction assay. We showed that protein corona constituents might alter the fate of nanoparticles in vivo, e.g., by subjecting them to phagocytosis. We demonstrated that composition of protein corona of nanoparticles is species-specific that leads to dissimilar transfection efficiency and should be considered while developing delivery systems for nucleic acids.

scholarly journals Internalisation and biological activity of nucleic acids delivering cell-penetrating peptide nanoparticles is controlled by the biomolecular corona

2021 ◽  
Author(s):  
Annely Lorents ◽  
Maria Maloverjan ◽  
Kärt Padari ◽  
Margus Pooga
Keyword(s):  
Nucleic Acid ◽  
Nucleic Acids ◽  
Transfection Efficiency ◽  
Protein Corona ◽  
Blood Stream ◽  
Cell Penetrating Peptides ◽  
Biological Efficiency ◽  
Cell Penetrating

Nucleic acid molecules can be transferred into cells to alter gene expression and, thus, alleviate certain pathological conditions. Cell-penetrating peptides (CPPs) are vectors that can be used for transfecting nucleic acids as well as many other compounds. CPPs associate nucleic acids non-covalently, forming stable nanoparticles and providing efficient transfection of cells in vitro. However, in vivo, expected efficiency is achieved only in rare cases. One of the reasons for this discrepancy is formation of protein corona around nanoparticles, once they are exposed to a biological environment, e.g. blood stream. In this study, we compared CPP-nucleic acid nanoparticles formed in the presence of bovine, murine and human serum. We used Western blot and mass-spectrometry to identify the major constituents of protein corona forming around nanoparticles, showing that proteins involved in transport, haemostasis and complement system are its major components. We investigated physical features of nanoparticles, and measured their biological efficiency in splice-correction assay. We showed that protein corona constituents might alter the fate of nanoparticles in vivo, e.g. by subjecting them to phagocytosis. We demonstrated that composition of protein corona of nanoparticles is species-specific that leads to dissimilar transfection efficiency and should be taken into account while developing delivery systems for nucleic acids.

scholarly journals Cell Penetrating Peptides Mediated DNA-Free Transfection in Arabidopsis and Chinese Cabbage

2021 ◽  
Author(s):  
Han Wu ◽  
Zhipeng Zhang ◽  
Kai Zhu ◽  
Fulai Ke ◽  
Fei Zhang ◽  
...  
Keyword(s):  
Fusion Protein ◽  
Genome Editing ◽  
Chinese Cabbage ◽  
Dna Sequences ◽  
Room Temperature ◽  
Genetically Modified Organisms ◽  
Transfection Efficiency ◽  
Cell Penetrating Peptides ◽  
Root Tips ◽  
Cell Penetrating

Abstract Background: The third-generation genome editing system CRISPR/Cas had shown strong application prospects in crop genetic improvement. However, this technology largely depends on genetic transformation. Public concerns on GMO (genetically modified organisms) safety, as well as related regulations, have restricted the application. Therefore, establishing DNA-free transfection system is important to promote CRISPR/Cas genome editing in agriculture.Results：In this paper, cell penetrating peptides fusion protein (CPP-mCherry) was found to be effective on DNA-free transfection. DNA sequences of nine tandem arginine (R9), one cysteine (cys), reporter mCherry and histidine label were sequentially constructed into pET 45B+ expression vector and transformed into Escherichia coli BL21(DE3) strain. CPP-mCherry fusion protein can be induced by 1mM IPTG for at least 1 hour in 28 °C. CPP-mCherry fusion protein can be obtained by 200W ultrasonication, then purified by Ni column and MWCO dialysis. The Arabidopsis thaliana root tips and leaves, as well as Chinese cabbage microspores and 3-week-old microspore embryo can be used as transfected recipient. Concentration can be selected between 10-100μg/ml and incubated overnight at room temperature. R9-cys-mCherry protein can be translocated into the nucleus of microspore. The transfection efficiency of root tips reached 100% and of microspore and MDE was 8.13% and 94.79%, respectively. Conclusions: Here, a CPP mediated DNA-free transfection system was built in dicots. These results lay a technical foundation of DNA-free genome editing.

scholarly journals Divalent metal ions boost effect of nucleic acids delivered by cell-penetrating peptides

2021 ◽  
Author(s):  
Maria Maloverjan ◽  
Kart Padari ◽  
Aare Abroi ◽  
Ana Rebane ◽  
Margus Pooga
Keyword(s):  
Nucleic Acids ◽  
Therapeutic Potential ◽  
Transfection Efficiency ◽  
Cell Penetrating Peptides ◽  
In Vivo Studies ◽  
Divalent Metal Ions ◽  
Reporter Protein ◽  
Cell Penetrating ◽  
Novel Strategy

Cell-penetrating peptides (CPPs) are promising tools for transfection of various substances, including nucleic acids, into cells. The aim of current work was to search for novel safe and effective approaches for enhancing transfection efficiency of nanoparticles formed of CPP and splice-correcting oligonucleotide (SCO) without increasing the concentration of peptide. We analyzed an effect of inclusion of calcium and magnesium ions into nanoparticles on CPP-mediated transfection in cell culture. We also studied the mechanism of such transfection as well as its efficiency, applicability in case of different cell lines, nucleic acid types and peptides, and possible limitations. We discovered a strong positive effect of these ions on transfection efficiency of SCO, that translated to enhanced synthesis of functional reporter protein. We observed significant changes in intracellular distribution and trafficking of nanoparticles formed with addition of the ions, without increasing cytotoxicity. We propose a novel strategy of preparing CPP-oligonucleotide nanoparticles with enhanced efficiency and, thus, higher therapeutic potential. Our discovery may be translated to primary cell cultures and, possibly, in vivo studies, in the aim to increase CPP-mediated transfection efficiency and likelihood of using CPPs in clinics.

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