Fusion Protein Reprogramming via Cell Permeable Peptide
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Solution Overview
Problem
Current methods for preparing induced pluripotent stem cells face challenges such as low efficiency and stability due to the use of viruses for gene transfer, which can be toxic and cause cytotoxicity, and existing cell permeable peptides derived from viruses raise safety concerns.
Innovation Solution
A fusion protein is developed by combining a reprogramming inducing factor with a low molecular weight protamine cell permeable peptide, which is used to effectively transport the protein into human dental pulp-derived stem cells, allowing for efficient reprogramming without the need for viral vectors, using a recombinant polynucleotide and vector system.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Productivity
If viral vectors are used for gene transfer to induce reprogramming, then reprogramming efficiency is improved, but safety and stability deteriorate due to cytotoxicity and potential cancer formation
Solution Approach 1:
The patent extracts and utilizes only the cell-permeable peptide component (TAT domain) from the viral protein structure, separating it from the harmful viral genetic material. This allows the beneficial cell penetration function to be retained while eliminating the cytotoxic and oncogenic risks associated with viral vectors.
Solution Approach 2:
The cell-permeable peptide acts as an intermediary carrier that facilitates the delivery of reprogramming factors into cells without using viral vectors. The peptide mediates cell entry through a non-viral mechanism, thereby maintaining safety while achieving efficient delivery.
2Ease of operation
If existing viral-derived cell permeable peptides are used for protein delivery, then cell permeability is improved, but safety deteriorates due to potential toxicity concerns
Solution Approach 1:
The patent extracts only the functional TAT domain (amino acids 48-82) from the full-length TAT protein, isolating the minimal sequence required for cell permeability. This truncated version maintains cell entry capability while reducing the molecular complexity and potential toxicity associated with the complete viral protein.
3Object-affected harmful factors
If liposome-based gene transfer is used, then cytotoxicity is reduced compared to electrical stimulation, but permeation efficiency deteriorates significantly
Solution Approach 1:
The cell-permeable peptide serves as a molecular mediator that directly interacts with the cell membrane to facilitate protein entry. Unlike liposomes that rely on endocytosis, the peptide acts as an active carrier that shuttles the reprogramming factors across the membrane barrier, achieving both high efficiency and low toxicity.
Applied Scientific Principles
This section explains which scientific principles are used to turn an abstract innovation direction into a practical engineering solution.
Function Achieved in This Case
This approach enables the high-efficiency and stable preparation of induced pluripotent stem cells by maximizing the effect of reprogramming inducing transcription factors, overcoming the limitations of existing viral peptide transporters and ensuring safety by avoiding viral vectors.
Implementation Method 1
A fusion protein is developed by combining a reprogramming inducing factor with a low molecular weight protamine cell permeable peptide, which is used to effectively transport the protein into human dental pulp-derived stem cells
Data Source
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AI summary
A method of preparing a reprogramming induced pluripotent stem cell from a human-derived somatic cell using a fusion protein in which a reprogramming inducing factor and cell permeable peptide (CPP) are fused, and a fusion protein in which a reprogramming inducing factor and a cell permeable peptide are fused are disclosed. According to the present invention, the induced pluripotent stem cell having high efficiency and high stability can be prepared by maximizing the effect of the reprogramming inducing transcription factor beyond the existing viral peptide transporter, in inducing the reprogramming of the somatic cell.