Nuclear Science and Techniques

《核技术》(英文版) ISSN 1001-8042 CN 31-1559/TL     2019 Impact factor 1.556

Nuclear Science and Techniques ›› 2017, Vol. 28 ›› Issue (1): 11 doi: 10.1007/s41365-016-0165-8

• NUCLEAR CHEMISTRY,RADIOCHEMISTRY,RADIOPHARMACEUTICALS AND NUCLEAR MEDICINE • Previous Articles     Next Articles

Expression and radiolabeling of Cas9 protein

Qing-Long Yan 1,2, Hua-Ting Kong 2, Kai Xia 2, Yu Zhang 2, Ali Aladlbahi 3, Ji-Ye Shi 4, Li-Hua Wang 2, Chun-Hai Fan 2, Yun Zhao 1, Ying Zhu2   

  1. 1 Key Laboratory of Bio-Resources and Eco-Environment, Ministry of Education, College of Life Sciences, Sichuan University, Chengdu 610064, China
    2 Division of Physical Biology and Bioimaging Center, CAS Key Laboratory of Interfacial Physics and Technology, Shanghai Institute of Applied Physics, Chinese Academy of Sciences, Jiading Campus, Shanghai 201800, China
    3 Chemistry Department, King Saud University, Riyadh 11451, Saudi Arabia
    4 UCB Pharma, Slough SL1 3WE, UK
  • Contact: Ying Zhu E-mail:zhuying@sinap.ac.cn
  • Supported by:

    This study was supported by the National Key Research and Development Program (No. 2016YFA0400902), the Ministry of Science and Technology of China (Nos. 2012CB825805, 2012CB932600), the National Natural Science Foundation of China (Nos. 11675251 and 11275251), Shanghai Rising-Star Program (No. 14QA1404400), Distinguished Scientist Fellowship Program of King Saud University, and the Youth Innovation Promotion Association of CAS (No. 2016236).

Qing-Long Yan, Hua-Ting Kong, Kai Xia, Yu Zhang, Ali Aladlbahi, Ji-Ye Shi, Li-Hua Wang, Chun-Hai Fan, Yun Zhao, Ying Zhu. Expression and radiolabeling of Cas9 protein.Nuclear Science and Techniques, 2017, 28(1): 11     doi: 10.1007/s41365-016-0165-8

Abstract:

As a robust platform for genome editing, CRISPR/Cas9 is currently being explored for engineering biology or therapeutics, yet means for quantitative detection of Cas9 proteins remain to be fully realized. Here, we expressed Cas9 proteins and developed a novel detection method that traced Cas9 based on radiolabeled iodine. Through optimizing the reaction conditions of reaction time, temperature and cycles, we obtained 125I-Cas9 of high labeling yield. The prepared 125I-Cas9 was stable in various media and preserved excellent genome editing efficiency. Thus, our strategy provides a convenient and efficient tool for further tracing biological behaviors of Cas9 proteins in living systems.

Key words: Cas9, Radiolabeling, 125I labeling yield, Stability