MACF1 Overexpression by Transfecting the 21 kbp Large Plasmid PEGFP-C1A-ACF7 Promotes Osteoblast Differentiation and Bone Formation

Yan Zhang, Chong Yin, Lifang Hu, Zhihao Chen, Fan Zhao, Dijie Li, Jianhua Ma, Xiaoli Ma, Peihong Su, Wuxia Qiu, Chaofei Yang, Pai Wang, Siyu Li, Ge ZHANG, Liping Wang*, Airong Qian, Cory J. Xian

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

12 Citations (Scopus)

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.

Original languageEnglish
Pages (from-to)259-270
Number of pages12
JournalHuman Gene Therapy
Volume29
Issue number2
DOIs
Publication statusPublished - Feb 2018

Scopus Subject Areas

  • Molecular Medicine
  • Molecular Biology
  • Genetics

User-Defined Keywords

  • bone formation
  • large plasmid
  • MACF1
  • osteoblast

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