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

doi:10.1007/s10529-016-2046-y

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

Pediatrics

Research output: Contribution to journal › Article › peer-review

6 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 language	English (US)
Pages (from-to)	893-900
Number of pages	8
Journal	Biotechnology Letters
Volume	38
Issue number	5
DOIs	https://doi.org/10.1007/s10529-016-2046-y
State	Published - May 1 2016

Funding

This work was supported by Grants from the Special Foundation for Science and Technology Key Plan of Guangdong Province (No. 2011A030400007); the Special Foundation for Science and Technology Key Plan of Guangzhou City (No. 2011Y1-00033-6); and the National Natural Science Foundation of China (No.30801184).

Keywords

Bone marrow stromal cells
Chondroitin sulfate proteoglycans
Chondroitinase ABC
Lentivirus
Secretion

ASJC Scopus subject areas

Applied Microbiology and Biotechnology
Bioengineering
Biotechnology

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10.1007/s10529-016-2046-y

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title = "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",

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.",

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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. / Guo, Yang; Klüppel, Michael; Tang, Hao et al.
In: Biotechnology Letters, Vol. 38, No. 5, 01.05.2016, p. 893-900.

Research output: Contribution to journal › Article › peer-review

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AU - Guo, Yang

AU - Klüppel, Michael

AU - Tang, Hao

AU - Tan, Sheng

AU - Zhang, Peidong

AU - Chen, Zhenzhou

PY - 2016/5/1

Y1 - 2016/5/1

N2 - 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.

AB - 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.

KW - Bone marrow stromal cells

KW - Chondroitin sulfate proteoglycans

KW - Chondroitinase ABC

KW - Lentivirus

KW - Secretion

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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

Abstract

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