Scholarlink

<p>The potential of siRNA to silence aberrantly expressed genes involved in a variety of diseases makes it a promising therapeutic approach in medical communities. However, poor cell membrane permeability and degradation through endo-lysosomal trapping present major obstacles to its clinical application, necessitating the need for an effective delivery vector. Several peptide sequences, known as cell-penetrating peptides (CPPs), have been reported to transport cell-impermeable cargoes across cellular membranes. In this study, we aimed to design a peptide-siRNA complex for efficient cellular delivery of anti-Smad7 siRNA (siSmad7) as a potential therapeutic approach for colorectal cancer. To achieve this, we carried out a comprehensive screening of peptide databases and identified a cationic peptide (GT) derived from the Oct-6 transcription factor, which had previously demonstrated high cytoplasmic accumulation. The GT-siSmad7 complexes were prepared at different peptide to siRNA molar ratios, and the optimal ratio was selected based on the highest siRNA binding efficiency (75%). Investigating the uptake mechanism by flow cytometry revealed the direct translocation of the peptide-siRNA complex across the cell membrane of HCT-116 cells, resulting in approximately 60% downregulation of Smad7 mRNA and about a 50% decrease in Smad7 protein levels, as determined by real-time PCR and western blot analysis, respectively. Reducing Smad7 levels induced apoptosis in nearly 60% of cancer cells. The GT peptide, derived from an endogenous transcription factor, demonstrated energy-independent uptake, and minimal toxicity, making it a promising and biocompatible delivery system for siRNA.</p> Graphical Abstract <p></p>

错误：搜索内容不能为空，请输入英文关键词
错误：关键词超出字数限制，请精简
高级检索

Efficient Transfection of siRNA with a Transcription Factor Oct-6-Derived Peptide

  • 期刊论文

  • 发表日期：

    2026年1月7日

查看全文
  • Elham Seyyednia,
  • Amir Baghbanzadeh,
  • Omid Rahbar Farzam,
  • Amir Ali Mokhtarzadeh,
  • Behzad Baradaran,
  • Javid Shahbazi Mojarrad,
  • Hadi Valizadeh

摘要

The potential of siRNA to silence aberrantly expressed genes involved in a variety of diseases makes it a promising therapeutic approach in medical communities. However, poor cell membrane permeability and degradation through endo-lysosomal trapping present major obstacles to its clinical application, necessitating the need for an effective delivery vector. Several peptide sequences, known as cell-penetrating peptides (CPPs), have been reported to transport cell-impermeable cargoes across cellular membranes. In this study, we aimed to design a peptide-siRNA complex for efficient cellular delivery of anti-Smad7 siRNA (siSmad7) as a potential therapeutic approach for colorectal cancer. To achieve this, we carried out a comprehensive screening of peptide databases and identified a cationic peptide (GT) derived from the Oct-6 transcription factor, which had previously demonstrated high cytoplasmic accumulation. The GT-siSmad7 complexes were prepared at different peptide to siRNA molar ratios, and the optimal ratio was selected based on the highest siRNA binding efficiency (75%). Investigating the uptake mechanism by flow cytometry revealed the direct translocation of the peptide-siRNA complex across the cell membrane of HCT-116 cells, resulting in approximately 60% downregulation of Smad7 mRNA and about a 50% decrease in Smad7 protein levels, as determined by real-time PCR and western blot analysis, respectively. Reducing Smad7 levels induced apoptosis in nearly 60% of cancer cells. The GT peptide, derived from an endogenous transcription factor, demonstrated energy-independent uptake, and minimal toxicity, making it a promising and biocompatible delivery system for siRNA.

Graphical Abstract