Biotechnology Letters

, Volume 39, Issue 10, pp 1471–1476 | Cite as

Genetically engineered cell lines for α1-antitrypsin expression

  • Qianqian Ji
  • Caiping Guo
  • Chen Xie
  • Yingdan Wu
  • Pei Zhang
  • Hui Li
  • Yongjun LuEmail author
Original Research Paper



To establish genetically modified cell lines that can produce functional α1-antitrypsin (AAT), by CRISPR/Cas9-assisted homologous recombination.


α1-Antitrypsin deficiency (AATD) is a monogenic heritable disease that often results in lungs and liver damage. Current augmentation therapy is expensive and in short of supply. To develop a safer and more effective therapeutic strategy for AATD, we integrated the AAT gene (SERPINA1, NG_008290.1) into the AAVS1 locus of human cell line HEK293T and assessed the safety and efficacy of CRISPR/Cas9 on producing potential therapeutic cell lines. Cell clones obtained had the AAT gene integrated at the AAVS1 locus and secreted approx. 0.04 g/l recombinant AAT into the medium. Moreover, the secreted AAT showed an inhibitory activity that is comparable to plasma AAT.


CRISPR/Cas9-mediated engineering of human cells is a promising alternative for generating isogenic cell lines with consistent AAT production. This work sheds new light on the generation of therapeutic liver stem cells for AATD.


AAVS1 locus α1-Antitrypsin deficiency CRISPR/Cas9 Homologous recombination 



This work was supported by China Postdoctoral Science Foundation (Grant No. 2015M582465).

Supporting information

Supplementary Table 1—Primer sequences.

Supplementary Figure 1—SDS-PAGE of AAT in puromycin-resistant HEK293T cells after transfection.

Compliance with ethical standards

Conflicts of interest

The authors have no conflict of interest to declare, and no competing financial interests exist.

Supplementary material

10529_2017_2391_MOESM1_ESM.docx (562 kb)
Supplementary material 1 (DOCX 562 kb)


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Copyright information

© Springer Science+Business Media B.V. 2017

Authors and Affiliations

  1. 1.School of Life SciencesSun Yat-Sen UniversityGuangzhouPeople’s Republic of China
  2. 2.Shenzhen Weiguang Biological Products Co., Ltd.ShenzhenPeople’s Republic of China
  3. 3.Guang ZhouPeople’s Republic of China

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