使用终端脱核样转移酶直接记录绑定信息到DNA编码的库中
Lukas A Schneider1, Basilius Sauter1, Koder Dagher1
1Department of Chemistry, University of Basel, 4056 Basel, Switzerland.
Journal of the American Chemical Society
|September 13, 2023
概括
终端脱核基转移酶 (TdT) 通过产生与结合亲和率成比例的多氨酸尾巴来实现DNA编码库 (DEL) 结合剂选择. 这种方法有效地识别了广泛的亲和度范围内的结合剂,提高了DEL选能力.
科学领域:
- 生物化学
- 分子生物学
- 药物发现
背景情况:
- 终端脱核基转移酶 (TdT) 是一种独特的DNA聚合酶.
- DNA编码库 (DEL) 是药物发现的强大工具.
- 传统的亲和选择方法在识别具有中等亲和力的结合剂方面存在局限性.
研究的目的:
- 开发一种使用TdT识别DEL的结合剂的新方法.
- 提高DEL选中的灵敏度和亲和度范围.
- 在DEL中创建一个稳定的分子记录.
主要方法:
- 目标蛋白与TdT融合在一起.
- 在dATP的存在下使用DEL化TdT融合蛋白.
- 在DNA结合剂上的多氨酸 (polyA) 尾部合成,尾部长度与结合亲和力相关.
- 使用磁性多晶dT25珠进行结合剂的丰富.
- 测序以识别富含的结合剂.
主要成果:
- 在DNA结合剂上成功生成了多A尾部.
- PolyA 尾巴的长度作为结合亲和力的直接测量.
- 鉴定出具有纳米分子到双位数微分子结合亲缘关系的配体.
- 这种方法在更广泛的亲和范围识别中优于经典亲和选择.
结论:
- 以TdT为媒介的多A尾部合成是一种简单有效的DEL结合剂选择方法.
- 与传统方法相比,该技术显著扩大了可检测的结合亲和度范围.
- 这种方法提供了一种强大而通用的工具,以加快使用DEL的药物发现.
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