@article{4537c54c6e9a4a2ca1ad4fa65178956c,
title = "MACF1 Overexpression by Transfecting the 21 kbp Large Plasmid PEGFP-C1A-ACF7 Promotes Osteoblast Differentiation and Bone Formation",
abstract = "Microtubule actin crosslinking factor 1 (MACF1) is a large spectraplakin protein known to have crucial roles in regulating cytoskeletal dynamics, cell migration, growth, and differentiation. However, its role and action mechanism in bone remain unclear. The present study investigated optimal conditions for effective transfection of the large plasmid PEGFP-C1A-ACF7 (∼21 kbp) containing full-length human MACF1 cDNA, as well as the potential role of MACF1 in bone formation. To enhance MACF1 expression, the plasmid was transfected into osteogenic cells by electroporation in vitro and into mouse calvaria with nanoparticles. Then, transfection efficiency, osteogenic marker expression, calvarial thickness, and bone formation were analyzed. Notably, MACF1 overexpression triggered a drastic increase in osteogenic gene expression, alkaline phosphatase activity, and matrix mineralization in vitro. Mouse calvarial thickness, mineral apposition rate, and osteogenic marker protein expression were significantly enhanced by local transfection. In addition, MACF1 overexpression promoted β-catenin expression and signaling. In conclusion, MACF1 overexpression by transfecting the large plasmid containing full-length MACF1 cDNA promotes osteoblast differentiation and bone formation via β-catenin signaling. Current data will provide useful experimental parameters for the transfection of large plasmids and a novel strategy based on promoting bone formation for prevention and therapy of bone disorders.",
keywords = "bone formation, large plasmid, MACF1, osteoblast",
author = "Yan Zhang and Chong Yin and Lifang Hu and Zhihao Chen and Fan Zhao and Dijie Li and Jianhua Ma and Xiaoli Ma and Peihong Su and Wuxia Qiu and Chaofei Yang and Pai Wang and Siyu Li and Ge Zhang and Liping Wang and Airong Qian and Xian, {Cory J.}",
note = "Funding Information: This work was supported by the National Natural Science Foundation of China (31570940, 31400725, and 81671928), China Postdoctoral Science Foundation (2015T81051), Shenzhen Science and Technology Project (JCYJ20160229174320053), and Fundamental Research Funds for the Central Universities (3102016ZY037). L.W. was supported by an Australian National Health and Medical Research Council grant (1094606) and the National Natural Science Foundation of China (81671928), and C.J.X. was supported by an Australian National Health and Medical Research Council Senior Research Fellowship (1042105) and the National Natural Science Foundation of China (81671928). Funding Information: This work was supported by the National Natural Science Foundation of China (31570940, 31400725, and 81671928), China Postdoctoral Science Foundation (2015T81051), Shenzhen Science and Technology Project ( JCYJ20160229174320053), and Fundamental Research Funds for the Central Universities (3102016ZY037). L.W. was supported by an Australian National Health and Medical Research Council grant (1094606) and the National Natural Science Foundation of China (81671928), and C.J.X. was supported by an Australian National Health and Medical Research Council Senior Research Fellowship (1042105) and the National Natural Science Foundation of China (81671928).",
year = "2018",
month = feb,
doi = "10.1089/hum.2017.153",
language = "English",
volume = "29",
pages = "259--270",
journal = "Human Gene Therapy",
issn = "1043-0342",
publisher = "Mary Ann Liebert Inc.",
number = "2",
}