[New option for gene-targeting vector construction Red/ET recombination].

Area of Science:

• Molecular Biology
• Genetics
• Gene Editing

Context:

• Gene function studies increasingly rely on gene knockout (KO) mouse models.
• Efficient construction of targeting vectors is critical for generating KO mice.
• Traditional methods for vector construction have low recombination efficiency.

Purpose:

• To develop an efficient method for constructing targeting vectors for new gene study.
• To utilize the Red/ET recombination DNA engineering platform for vector construction.
• To create targeting vectors for the novel gene Resp18.

Summary:

• Red/ET recombineering was employed to construct Resp18 targeting vectors through two homologous recombination processes.
• This method facilitates gene modification via acquisition, insertion, fusion, or replacement using small fragments.
• Three distinct Resp18 targeting vectors were successfully generated without mutations and with longer homology regions.

Impact:

• Provides a new and reliable alternative for constructing gene targeting vectors.
• Enhances the efficiency and accuracy of generating gene knockout mouse models.
• Facilitates accelerated research into the functions of newly discovered genes.