Title: Development novel and efficient peptides for gene delivery
Abstract:
The most important problem in the gene therapy is gene delivery. Cell-penetrating peptides (CPPs) have emerged as versatile tools for delivering various cargoes into cells. However, the exact mechanism of cellular uptake by CPPs remains elusive. This study aimed to explore the efficacy of new peptides, a specific class of CPP-nucleic acid binding peptide hybrids as gene delivery agents. A novel, highly efficient, cost- effective method was developed using these peptides. Eight peptides (named TM1 to TM8), composed of a nucleic acid binding part and a hydrophobic part of different cell-penetrating peptides, were synthesized, and tested for their ability to facilitate intracellular delivery of plasmid DNA. The results demonstrated that TM3 peptide exhibited the best performance among all TM peptides, in delivering the gene of interest to the nucleus. This study highlights the potential of TM peptides, specifically TM3, as a novel gene delivery vector in the field of gene therapy.
Cell-penetrating peptides (CPPs) have garnered significant attention due to their ability to traverse cellular membranes and deliver diverse cargoes, including drugs, nucleic acids, and proteins, into cells. This unique property of CPPs has positioned them as promising tools for drug delivery and various biomedical applications. While CPPs are known to penetrate cells through mechanisms such as endocytosis, direct translocation, or a combination thereof, the precise mechanism of cellular uptake by CPPs remains to be clarified (1). Currently liposomes, vesicles composed of phospholipids capable of encapsulating hydrophilic and hydrophobic molecules, are commonly used for gene delivery (2). This study presents a novel method characterized by high efficacy, low toxicity, and cost-effectiveness agent compared to liposomes, as a gene delivery tool.
Tools and Method: Eight TM peptides were designed and synthesized using a peptide synthesizer. These peptides incorporated a short positively charged segment derived from nucleic acid binding peptide and a hydrophobic portion from different cell-penetrating peptides. The synthesis of TM peptides was followed by incubation with different dilutions of pcDNA-GFP (which expresses green fluorescent protein in the cell), at room temperature. The resulting hybrid peptide-pcDNA-GFP complex was then delivered into HeLa cells.
Results: The investigation revealed that the utilization of TM peptides facilitated fast and efficient expression of GFP which indicated successful delivery of pcDNA-GFP into HeLa cells, achieving cellular entry efficiency as high as 70-80%. Among the tested TM peptides, TM3 exhibited superior performance compared to other TM variants, as confirmed by fluorescence microscopy assays measuring total cellular uptake and cytosolic delivery.
Discussion: In conclusion, this study highlights the potential of TM3 peptide as a novel gene delivery agent in gene therapy. The efficient intracellular delivery and nuclear-targeting capabilities of TM3 make it an attractive candidate for targeted gene delivery applications. The development of TM peptides as gene-delivery vectors offers promising opportunities for advancing gene therapy and treating various diseases. Further optimization and investigations are warranted to fully exploit the potential of TM peptides in gene delivery applications. TM peptides may be an efficient tool of gene therapy.
Audience Take Away Notes:
- Audience members will gain insights into the efficacy of novel peptides, specifically TM3, for gene delivery in the context of gene therapy. They'll understand the methodology behind synthesizing and testing these peptides and how they compare to traditional liposomes in terms of efficacy, toxicity, and cost-effectiveness
- Professionals in the field of gene therapy and drug delivery will benefit from this research by having access to a potential new tool for more efficient and targeted gene delivery. This could lead to advancements in their work, offering improved treatment options for various diseases
- Yes, this research provides valuable insights and methodologies that other faculty members could utilize to expand their own research in gene therapy, drug delivery, or related fields. Additionally, it could serve as educational material for teaching students about advanced techniques in biomedicine
- Absolutely. By offering a highly efficient and cost-effective method forgene delivery compared to traditional liposomes, this research simplifies the process and potentially reduces costs associated with gene therapy. This could make the job of designing gene delivery systems more efficient and accessible
- Yes, the study demonstrates the superior performance of TM3 peptide in delivering genes to the nucleus, which can enhance the accuracy and efficacy of gene delivery systems. Additionally, it provides new information about the potential of these peptides as gene delivery vectors, which can assist in designing more effective gene therapy treatments
