TY - JOUR
T1 - 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
AU - Guo, Yang
AU - Klüppel, Michael
AU - Tang, Hao
AU - Tan, Sheng
AU - Zhang, Peidong
AU - Chen, Zhenzhou
N1 - Funding Information:
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).
Publisher Copyright:
© 2016, Springer Science+Business Media Dordrecht.
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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U2 - 10.1007/s10529-016-2046-y
DO - 10.1007/s10529-016-2046-y
M3 - Article
C2 - 26910777
AN - SCOPUS:84959137557
SN - 0141-5492
VL - 38
SP - 893
EP - 900
JO - Biotechnology Letters
JF - Biotechnology Letters
IS - 5
ER -