Zhang Kaili, Wang Yi, Li Yifan, Jiang Shuze, Chen Yongzi, Chen Min, Su Weijun, Liu Liren, Li Shuai
Department of Molecular Pharmacology, Tianjin Medical University Cancer Institute and Hospital; National Clinical Research Center for Cancer; Key Laboratory of Cancer Prevention and Therapy, Tianjin, China.
Tianjin's Clinical Research Center for Cancer, Tianjin, China.
Methods Mol Biol. 2025;2934:293-303. doi: 10.1007/978-1-0716-4578-9_19.
The construction of DNA-encoded peptide libraries in mammalian cells is an evolving field that combines the advantages of mammalian cell expression systems with the vast diversity of DNA-encoded libraries. This enables high-throughput screening and identification of functional peptides in a mammalian cell context, aiding in the elucidation of disease-related proteins and the development of novel peptide drugs. We have developed a strategy for constructing dsDNA-based AND-gate (DBAG), which utilizes dsDNAs with NNK degenerate codons at the ends to implement AND-gate genetic circuits and generate peptide libraries in mammalian cells. We provide a detailed description of the library preparation, transfection, and analysis methods.
在哺乳动物细胞中构建DNA编码肽库是一个不断发展的领域，它将哺乳动物细胞表达系统的优势与DNA编码文库的巨大多样性结合在一起。这使得在哺乳动物细胞环境中能够进行高通量筛选和功能性肽的鉴定，有助于阐明疾病相关蛋白并开发新型肽药物。我们已经开发了一种构建基于双链DNA的与门（DBAG）的策略，该策略利用末端带有NNK简并密码子的双链DNA来实现与门遗传电路，并在哺乳动物细胞中生成肽库。我们详细描述了文库制备、转染和分析方法。
