Lentivirus-mediated transfection of chondroitinase ABC gene without the bacterial leader sequence enables long-term secretion of functional chondroitinase ABC in human bone marrow stromal cells

Yang Guo, Michael Klüppel, Hao Tang, Sheng Tan, Peidong Zhang, Zhenzhou Chen*

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

3 Scopus citations

Abstract

Objective: To test the feasibility of secretion of functional chondroitinase ABC (ChABC), a bacterial enzyme that promotes axonal regeneration after spinal cord injury, from human bone marrow stromal cells (hBMSCs). Results: A lentiviral-expression vector, Lenti6.3-ChABC-3F, carrying the ChABC-3F gene without the bacterial leader sequence (aa 1-24) was constructed. Transfection of these Lenti6.3-ChABC-3F lentivirus led to stable expression in and secrection of ChABC proteins from hBMSCs for at least ten passages in culture in vitro, which was demonstrated by QRT-PCR, immunostaining, western blotting and ELISA. Moreover, the secreted ChABC proteins exhibited similar functional activity as the commercially-available ChABC. Conclusions: The lentivirus-mediated transfection of chondroitinase ABC gene without the bacterial leader sequence induced substantial long-term secretion of functional ChABC in hBMSCs.

Original languageEnglish (US)
Pages (from-to)893-900
Number of pages8
JournalBiotechnology Letters
Volume38
Issue number5
DOIs
StatePublished - May 1 2016

Keywords

  • Bone marrow stromal cells
  • Chondroitin sulfate proteoglycans
  • Chondroitinase ABC
  • Lentivirus
  • Secretion

ASJC Scopus subject areas

  • Biotechnology
  • Bioengineering
  • Applied Microbiology and Biotechnology

Fingerprint Dive into the research topics of 'Lentivirus-mediated transfection of chondroitinase ABC gene without the bacterial leader sequence enables long-term secretion of functional chondroitinase ABC in human bone marrow stromal cells'. Together they form a unique fingerprint.

Cite this