Hydrophobic and electrostatic interactions between cell penetrating peptides and plasmid DNA are important for stable non-covalent complexation and intracellular delivery

Cell penetrating peptides are useful tools for intracellular delivery of nucleic acids. Delivery of plasmid DNA, a large nucleic acid, poses a challenge for peptide mediated transport. The paper investigates and compares efficacy of five novel peptide designs for complexation of plasmid DNA and subs...

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Bibliographic Details
Published in:Journal of peptide science Vol. 22; no. 10; pp. 647 - 659
Main Authors: Upadhya, Archana, Sangave, Preeti C.
Format: Journal Article
Language:English
Published: England Blackwell Publishing Ltd 01.10.2016
Wiley Subscription Services, Inc
Subjects:
Alkaline Phosphatase - genetics
Alkaline Phosphatase - secretion
Amino Acid Sequence
Animals
cell penetrating peptides
Cell Survival - drug effects
Cell-Penetrating Peptides - chemistry
Cell-Penetrating Peptides - metabolism
Cell-Penetrating Peptides - pharmacology
Cytotoxicity
Deoxyribonuclease I - chemistry
Deoxyribonucleic acid
DNA
DNA - chemistry
DNA - metabolism
Erythrocytes - drug effects
gene delivery
Gene Expression
Genes, Reporter
HeLa Cells
Hemolysis - drug effects
Humans
Hydrophobic and Hydrophilic Interactions
hydrophobic interaction
membrane translocating sequence
Mice
NIH 3T3 Cells
noncovalent complexation
Nucleic acids
Peptides
Plasmids - chemistry
Plasmids - metabolism
Protein Binding
Rats
Static Electricity
stearylated octa-arginine
Transfection - methods
gene delivery
cell penetrating peptides
stearylated octa-arginine
hydrophobic interaction
noncovalent complexation
membrane translocating sequence
ISSN:1075-2617, 1099-1387, 1099-1387
Online Access:Get full text
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Summary:Cell penetrating peptides are useful tools for intracellular delivery of nucleic acids. Delivery of plasmid DNA, a large nucleic acid, poses a challenge for peptide mediated transport. The paper investigates and compares efficacy of five novel peptide designs for complexation of plasmid DNA and subsequent delivery into cells. The peptides were designed to contain reported DNA condensing agents and basic cell penetrating sequences, octa‐arginine (R8) and CHK6HC coupled to cell penetration accelerating peptides such as Bax inhibitory mutant peptide (KLPVM) and a peptide derived from the Kaposi fibroblast growth factor (kFGF) membrane translocating sequence. A tryptophan rich peptide, an analogue of Pep‐3, flanked with CH3 on either ends was also a part of the study. The peptides were analysed for plasmid DNA complexation, protection of peptide–plasmid DNA complexes against DNase I, serum components and competitive ligands by simple agarose gel electrophoresis techniques. Hemolysis of rat red blood corpuscles (RBCs) in the presence of the peptides was used as a measure of peptide cytotoxicity. Plasmid DNA delivery through the designed peptides was evaluated in two cell lines, human cervical cancer cell line (HeLa) and (NIH/3 T3) mouse embryonic fibroblasts via expression of the secreted alkaline phosphatase (SEAP) reporter gene. The importance of hydrophobic sequences in addition to cationic sequences in peptides for non‐covalent plasmid DNA complexation and delivery has been illustrated. An alternative to the employment of fatty acid moieties for enhanced gene transfer has been proposed. Comparison of peptides for plasmid DNA complexation and delivery of peptide–plasmid DNA complexes to cells estimated by expression of a reporter gene, SEAP. Copyright © 2016 European Peptide Society and John Wiley & Sons, Ltd. Hydrophobic and electrostatic interactions between cell penetrating peptides and plasmid DNA are important for stable non‐covalent complexation and intracellular delivery. Comparison of peptides for plasmid DNA complexation and delivery of peptide–plasmid DNA complexes to cells estimated by expression of a reporter gene, SEAP.
Bibliography:ark:/67375/WNG-VK6R646X-S
istex:0B2CAB3A2617D90B8A2960B7F1299940425F0675
ArticleID:PSC2927
ObjectType-Article-1
SourceType-Scholarly Journals-1
ObjectType-Feature-2
content type line 14
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ISSN:1075-2617
1099-1387
1099-1387
DOI:10.1002/psc.2927