Fusion Polypeptide Gene Regulation via Cell-Penetrating Peptides
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Solution Overview
Problem
Current methods for regulating gene transcription in cells require complex procedures such as cell transfection or electroporation, which are invasive and inefficient.
Innovation Solution
A novel fusion polypeptide comprising a cell-penetrating peptide, a DNA-binding polypeptide, and a transcriptional activator, specifically designed to bind to target nucleotide sequences, allowing for the activation of gene transcription by simple addition to a cell culture medium.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If conventional transfection or electroporation methods are used to deliver transcriptional regulators into cells, then gene transcription regulation can be achieved, but the procedures become complex and invasive
Solution Approach 1:
The invention merges the cell-penetrating function, DNA-binding function, and transcriptional activation function into a single fusion polypeptide molecule. This integration eliminates the need for separate delivery systems and complex transfection procedures, allowing the fusion polypeptide to enter cells autonomously and directly regulate target gene transcription through simple addition to culture medium.
Solution Approach 2:
The fusion polypeptide possesses multiple functions simultaneously: the cell-penetrating peptide enables autonomous cell entry, the DNA-binding polypeptide provides specific target recognition, and the transcriptional activator domain executes gene regulation. This multi-functionality in a single molecule simplifies the overall system and eliminates the need for multiple separate components.
2Reliability
If conventional transfection methods are used, then transcriptional regulators can be delivered into cells, but the process becomes time-consuming and inefficient
Solution Approach 1:
The cell-penetrating peptide component of the fusion polypeptide enables autonomous cell entry without requiring external delivery assistance. The fusion polypeptide self-delivers into target cells through its inherent cell-penetrating capability, eliminating the need for time-consuming transfection or electroporation procedures and significantly improving delivery efficiency.
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
The fusion polypeptide effectively increases the expression of target genes, promoting cell growth and gene regulation without the need for invasive transfection methods, as demonstrated by increased mRNA and RNA levels and enhanced cell proliferation.
Implementation Method 1
Cell-penetrating peptides are polypeptides having the function of penetrating a cell membrane and moving to the inside of the cell
Implementation Method 2
a DNA-binding polypeptide that binds to the nucleotide sequence represented by SEQ ID NO: 49
Implementation Method 3
a transcriptional activator... increases the expression of a target gene when used with gRNA
Data Source
AI summary
It is intended to provide a fusion polypeptide that regulates the transcription of a target gene.The present inventors have provided a fusion polypeptide comprising: a cell-penetrating peptide; a DNA-binding polypeptide; and a transcriptional regulator.