Department of Pediatrics, Washington University School of Medicine, St. Louis, MO, USA.
Methods Mol Biol. 2022;2444:69-80. doi: 10.1007/978-1-0716-2063-2_5.
Development of B cells requires the programmed generation and repair of double-stranded DNA breaks in antigen receptor genes. Investigation of the cellular responses to these DNA breaks has established important insights into B cell development and, more broadly, has provided fundamental advances into the molecular mechanisms of DNA damage response pathways. Abelson transformed pre-B cell lines and primary pre-B cell cultures are malleable experimental systems with diverse applications for studying DNA damage responses. This chapter describes methods for generating these cellular systems, inducing and quantifying DSBs, and assessing DNA damage programs.
B 细胞的发育需要抗原受体基因中双链 DNA 断裂的程序性产生和修复。对这些 DNA 断裂的细胞反应的研究为 B 细胞发育提供了重要的见解，更广泛地说，为 DNA 损伤反应途径的分子机制提供了重要的进展。艾伯森转化的前 B 细胞系和原代前 B 细胞培养物是具有多种应用的可塑实验系统，可用于研究 DNA 损伤反应。本章介绍了生成这些细胞系统、诱导和量化 DSB 以及评估 DNA 损伤程序的方法。
