2. Academic Validation
  3. Generation of an Akaluc knock-in human embryonic stem cell reporter line using CRISPR-Cas9 technology

Generation of an Akaluc knock-in human embryonic stem cell reporter line using CRISPR-Cas9 technology

  • Stem Cell Res. 2021 Oct:56:102532. doi: 10.1016/j.scr.2021.102532.
Min Zhou 1,  Qi Xing 2,  Di Zhang 2,  Cong Zhang 2,  Yanqi Zhang 2,  Jingyuan Zhang 2,  Yongli Shan 3
Affiliations

Affiliations

  • 1 CAS Key Laboratory of Regenerative Biology, Guangdong Provincial Key Laboratory of Stem Cell and Regenerative Medicine, South China Institute for Stem Cell Biology and Regenerative Medicine, Guangzhou Institutes of Biomedicine and Health, Chinese Academy of Sciences, Guangzhou 510530, China.
  • 2 CAS Key Laboratory of Regenerative Biology, Guangdong Provincial Key Laboratory of Stem Cell and Regenerative Medicine, South China Institute for Stem Cell Biology and Regenerative Medicine, Guangzhou Institutes of Biomedicine and Health, Chinese Academy of Sciences, Guangzhou 510530, China; University of Chinese Academy of Sciences, Beijing 100049, China.
  • 3 CAS Key Laboratory of Regenerative Biology, Guangdong Provincial Key Laboratory of Stem Cell and Regenerative Medicine, South China Institute for Stem Cell Biology and Regenerative Medicine, Guangzhou Institutes of Biomedicine and Health, Chinese Academy of Sciences, Guangzhou 510530, China. Electronic address: shan_yongli@gibh.ac.cn.
PMID: 34509917 DOI: 10.1016/j.scr.2021.102532
Abstract

Akaluc, an enzyme engineered from luciferase, provides a potential powerful tool for tracing transplanted cells in vivo because of its near-infrared emission light. To enable evaluation of its potency, we inserted the Akaluc gene at AAVS1 locus using CRISPR/Cas9 technology and generated a clonal human embryonic stem stable cell line (Named H1-AAVS1-EF1α-Akaluc-KI or AkalucHES). AkalucHES could efficiently express Akaluc and were traced easily in vivo. We verified that AkalucHES expressed the pluripotency markers and showed normal stem cell morphology. Furthermore, AkalucHES maitains normal karyotype and is able to differentiate toward three germ-layer in vivo. So the Akaluc is effective for tracing transplanted cells in vivo.

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