Efficient construction of a stable linear gene based on a TNA loop modified primer pair for gene delivery

Area of Science:

• Molecular Biology
• Biotechnology
• Genetic Engineering

Background:

• Gene expression systems are crucial for genetic engineering and therapeutic applications.
• Current systems face challenges with stability and efficient delivery in eukaryotic cells.
• Developing robust nucleic acid structures is essential for overcoming these limitations.

Purpose of the Study:

• To construct a novel, terminal-closed linear gene with enhanced stability.
• To utilize α-l-threose nucleic acid (TNA) modified primers for gene synthesis.
• To evaluate the potential of this construct as an efficient gene expression system in eukaryotic cells.

Main Methods:

• Polymerase chain reaction (PCR) was employed for gene construction.
• α-l-threose nucleic acid (TNA) loop modified primer pairs were designed and synthesized.
• The constructed linear gene's exonuclease resistance and serum stability were assessed.
• Eukaryotic cell systems were used to evaluate gene expression efficiency.

Main Results:

• A terminal-closed linear gene construct was successfully synthesized.
• The gene exhibited significant exonuclease resistance and enhanced serum stability.
• The TNA-modified primer approach facilitated efficient gene expression in eukaryotic cells.
• The developed system demonstrated potential for effective gene delivery.

Conclusions:

• The novel terminal-closed linear gene construct offers superior stability and resistance.
• The use of TNA-modified primers is a viable strategy for enhancing gene expression systems.
• This engineered gene represents a promising tool for advanced gene delivery and expression in eukaryotic systems.