CRISPR/Cas9-mediated gene editing in human zygotes using Cas9 protein

Original Article

DOI: 10.1007/s00438-017-1299-z

Cite this article as:
Tang, L., Zeng, Y., Du, H. et al. Mol Genet Genomics (2017). doi:10.1007/s00438-017-1299-z


Previous works using human tripronuclear zygotes suggested that the clustered regularly interspaced short palindromic repeat (CRISPR)/Cas9 system could be a tool in correcting disease-causing mutations. However, whether this system was applicable in normal human (dual pronuclear, 2PN) zygotes was unclear. Here we demonstrate that CRISPR/Cas9 is also effective as a gene-editing tool in human 2PN zygotes. By injection of Cas9 protein complexed with the appropriate sgRNAs and homology donors into one-cell human embryos, we demonstrated efficient homologous recombination-mediated correction of point mutations in HBB and G6PD. However, our results also reveal limitations of this correction procedure and highlight the need for further research.


CRISPR/Cas9 Homology-directed repair (HDR) Cas9 protein Gene modification Human zygotes 

Supplementary material

438_2017_1299_MOESM1_ESM.docx (223 kb)
Supplementary material 1 (DOCX 222 KB)

Copyright information

© Springer-Verlag Berlin Heidelberg 2017

Authors and Affiliations

  1. 1.State Key Laboratory of Proteomics, Beijing Proteome Research CenterBeijing Institute of Radiation MedicineBeijingChina
  2. 2.National Center for Protein Sciences BeijingBeijingChina
  3. 3.Center for Reproductive Medicine, The Third Affiliated HospitalGuangzhou Medical UniversityGuangzhouChina
  4. 4.National Center for International Research of Biological Targeting Diagnosis and Therapy, Collaborative Innovation Center for Targeting Tumor Diagnosis and TherapyGuangxi Medical UniversityGuangxiChina
  5. 5.Houston Fertility InstituteHoustonUSA
  6. 6.Department of CardiologyBayi Hospital Affiliated Nanjing University of Chineses MedicineNanjingChina