ABSTRACT
Gene-based therapy largely relies on the vector type that allows a selective and efficient transfection into the target cells with maximum efficacy and minimal toxicity. Although, genes delivered utilizing modified viruses transfect efficiently and precisely, these vectors can cause severe immunological responses and are potentially carcinogenic. A promising method of overcoming this limitation is the use of non-viral vectors, including cationic lipids, polymers, dendrimers, and peptides, which offer potential routes for compacting DNA for targeted delivery. Although non-viral vectors exhibit reduced transfection efficiency compared to their viral counterpart, their superior biocompatibility, non-immunogenicity and potential for large-scale production make them increasingly attractive for modern therapy. There has been a great deal of interest in the development of biomimetic chimeric peptides. Biomimetic chimeric peptides contain different motifs for gene translocation into the nucleus of the desired cells. They have motifs for gene targeting into the desired cell, condense DNA into nanosize particles, translocate the gene into the nucleus and enhance the release of the particle into the cytoplasm. These carriers were developed in recent years. This review highlights the stepwise development of the biomimetic chimeric peptides currently being used in gene delivery.
Subject(s)
Biomimetic Materials/chemistry , DNA/chemistry , Gene Transfer Techniques , Peptides/chemistry , Amino Acid Motifs , Animals , Biomimetic Materials/pharmacology , DNA/pharmacology , Humans , Peptides/pharmacologyABSTRACT
OBJECTIVES: Fatigue is a common problem in modern-day life. The aim of this study was to evaluate the effect of Delphinium denudatum (Jadwar) on fatigue. METHODS: This study was a randomized double-blind placebo-controlled clinical trial between healthy normal university students. In each group, participants were given one capsule of either WEACURE® (containing 500â¯mg of Jadwar root powder) or placebo for 15 consecutive days. Multidimensional Fatigue Inventory (MFI) questionnaire was used before and after the intervention to evaluate different aspects of fatigue. RESULTS: A total number of 64 participants completed the study. Data analysis showed decrease in the scores of all five domains of fatigue in Jadwar group (13.31⯱â¯3.05-7.75⯱â¯2.66, 12.31⯱â¯3.55-7.63⯱â¯2.62, 12.22⯱â¯4.26-6.97⯱â¯2.06, 11.56⯱â¯4.21 to 7.28⯱â¯2.37, 12.91⯱â¯3.09-7.34⯱â¯2.13 in general fatigue, physical fatigue, reduced activity, reduced motivation, and mental fatigue domains, respectively) which was statistically significant (P value<0.0001). This situation was significantly superior to the placebo group. Prescribed dosage of WEACURE® capsule was well tolerated. CONCLUSION: As a complementary tonic agent, Jadwar have a potential to reduce fatigue in normal population. However, objective evaluation of its anti-fatigue effect should be further evaluated.
Subject(s)
Delphinium/chemistry , Fatigue/drug therapy , Plant Preparations/therapeutic use , Adult , Double-Blind Method , Female , Humans , Male , Surveys and Questionnaires , Treatment Outcome , Young AdultABSTRACT
Despite the initial successes of gene delivery applications, they faced on several intrinsic drawbacks including toxicity and immunogenicity. Therefore, alternative gene-delivery systems derived from recombinant peptides have emerged and is rapidly developing. Human epidermal growth factor receptor-3 (HER3) shows high activity in tumor resistance to anti-human epidermal growth factor receptor 2 (HER2) therapies. In this study, an affibody molecule against HER3 is conjugated to a biomimetic peptide RALA (an amphipathic and cationic peptide enriched with arginine) and the ability of the fusion vector for targeting HER3 and afterward delivering specific genes in breast cancer cells is evaluated. The results demonstrate that the biopolymeric platform, which contains an affibody-conjugated RALA peptide, can effectively condense DNA into nanoparticles and target the overexpressed HER3 receptors in breast cancer cells and transfer specific genes. The use of such a recombinant biopolymer may pave the way for the development of sensitive and effective diagnostic and treatment tool for breast cancer.
Subject(s)
Breast Neoplasms/therapy , Gene Transfer Techniques , Genetic Therapy/methods , Peptides/genetics , Receptor, ErbB-3/genetics , Recombinant Fusion Proteins/genetics , Amino Acid Sequence , Breast Neoplasms/genetics , Cell Line, Tumor , Cloning, Molecular , Female , Gene Expression Regulation, Neoplastic , HEK293 Cells , Humans , MCF-7 Cells , Nanoconjugates/chemistry , Nanotechnology/methods , Peptides/chemistry , Receptor, ErbB-3/chemistry , Receptor, ErbB-3/metabolism , Recombinant Fusion Proteins/chemistry , Recombinant Fusion Proteins/isolation & purification , Recombinant Fusion Proteins/metabolismABSTRACT
Several tissue specific non-viral carriers have been developed for gene delivery purposes. However, the inability to escape endosomes, undermines the efficacy of these carriers. Researchers inspired by HIV and influenza virus, have randomly used Gp41 and H5WYG fusogenic peptides in several gene delivery systems without any rational preference. Here for the first time, we have genetically engineered two Nano-biomimetic carriers composed of either HWYG (HNH) or Gp41 (GNH) that precisely provide identical conditions for the study and evaluation of these fusogenic peptides. The luciferase assay demonstrated a two-fold higher transfection efficiency of HNH compared to GNH. These nanocarriers also displayed equivalent properties in terms of DNA binding ability and DNA protection against serum nucleases and formed similar nanoparticles in terms of surface charge and size. Interestingly, hemolysis and cellular analysis demonstrated both of nanoparticles internalized into cells in similar rate and escaped from endosome with different efficiency. Furthermore, the structural analysis revealed the mechanisms responsible for the superior endosomal escaping capability of H5WYG. In conclusion, this study describes the rationale for using H5WYG peptide to deliver nucleic acids and suggests that using nano-biomimetic carriers to screen different endosomal release peptides, improves gene delivery significantly.
Subject(s)
Biomimetic Materials/chemistry , Gene Transfer Techniques , Intracellular Space/metabolism , Nanoparticles/chemistry , Amino Acid Sequence , Cell Death , DNA/metabolism , Drug Carriers/chemistry , Dynamic Light Scattering , HEK293 Cells , Hemolysis , Humans , Luciferases/metabolism , Particle Size , Peptides/chemistry , Serum/metabolism , TransfectionABSTRACT
Chimeric polymers are known as suitable carriers for gene delivery. Certain properties are critical for a polymer to be used as a gene delivery vector. A new polymer was designed for the targeted delivery of genes into breast cancer cell lines, based on MPG peptide. It is composed of different functional domains, including HIV gp41, nuclear localization sequence of SV40 T-antigen, two C-terminus repeats of histone H1, and the scFv of anti-HER2 antibody. The results demonstrated that the vector can effectively condense plasmid DNA into nanoparticles with an average size of 250nm. Moreover, fusion of the scFv portion to the carrier brought about the specific recognition of HER2. Overall, the transfection efficiency of the vector demonstrated that it could deliver the desired gene into BT-474 HER2-positive breast cancer cells.
Subject(s)
Breast Neoplasms/genetics , Breast Neoplasms/therapy , Peptides/chemistry , Peptides/genetics , Receptor, ErbB-2/genetics , Single-Chain Antibodies/metabolism , Breast Neoplasms/metabolism , Cell Line, Tumor , Female , Gene Transfer Techniques , Genetic Therapy/methods , Genetic Vectors/genetics , Humans , Nanoparticles/chemistry , Particle Size , Plasmids/genetics , Polymers/chemistry , Transfection/methodsABSTRACT
Bioluminescence reaction, which uses luciferin, Mg(2+)-ATP and molecular oxygen to yield an electronically excited oxyluciferin, is carried out by the luciferase and emit visible light. One of the most promising applications of firefly luciferase is biosensors. In order to develop an apoptosis biosensor based on caspase 3/7, we have generated 3 forms of circularly permuted variants of Photinus pyralis firefly luciferase and a relatively good tolerance toward disruption of the polypeptide chain by introduction of new termini were found. Two forms of circular permuted luciferases showed significant activity enhancement in comparison with control after exposure to caspase 3. Moreover, the effect of circular permutation and also the length of inserted peptide (caspase 3/7 recognition sites) in structure of firefly luciferase were analyzed using circular dichroism and fluorescence spectroscopy.