Expression of bioactive recombinant human fibroblast growth factor 9 in oil bodies of Arabidopsis thaliana

Yi, Shanyong; Yang, Jing; Huang, Jian; Guan, Lili; Du, Lina; Guo, Yongxin; Zhai, Feng; Wang, Yanfang; Lü, Zhou; Wang, Liyong; Li, Haiyan; Li, Xiaokun; Jiang, Chao

doi:10.1016/j.pep.2015.08.006

Cited by 18 publications

(16 citation statements)

References 30 publications

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“…The rhFGF9 expression vector was constructed and showed as previously report [ 8 ]. And clones carrying the desired expression vector were identified by PCR to evaluate insert size and verified by restriction enzyme digestion.…”

Section: Resultsmentioning

confidence: 99%

“…The pOTBar plasmid was used for the construction of rhFGF9 expression vector and the same method as previous report [ 8 ]. The Oleosin and rhFGF9 primers as following: oleosin forward, 5′- CCATGGCGGATACAGCTAGAGGAACC-3′; oleosin reverse, 5′- CTCTCCCAAAGGAGCCATAGTAGTGTGCTGGC-3′; rhFGF9 forward, 5′- GCCAGCACACTACTATGGCTCCTTTGGGAGAG-3′; rhFGF9 reverse, 5′- CCCAAGCTTAAGATTGAGAAAGGATATCCTTGT-3′.…”

Section: Methodsmentioning

confidence: 99%

“…It is an indispensable growth factor for human development. FGF9 contributes to bone development and repair, angiogenesis, embryonic development, cell apoptosis, nerve regeneration, and hair follicle regeneration [ 8 – 10 ]. In addition, FGF9 participates in the development of heart, brain, kidney, muscle, joint and other tissues [ 11 ].…”

Section: Introductionmentioning

confidence: 99%

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Oil body bound oleosin-rhFGF9 fusion protein expressed in safflower (Carthamus tinctorius L.) stimulates hair growth and wound healing in mice

Cai

Wen

et al. 2018

BMC Biotechnol

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BackgroundFibroblast growth factor 9 (FGF9) is a heparin-binding growth factor, secreted by both mesothelial and epithelial cells, which participates in hair follicle regeneration, wound healing, and bone development. A suitable source of recombinant human FGF9 (rhFGF9) is needed for research into potential clinical applications. We present that expression of oleosin-rhFGF9 fusion protein in safflower (Carthamus tinctorius L.) seeds stimulates hair growth and wound healing.ResultsThe oleosin-rhFGF9 expressed in safflower seeds, in which it localizes to the surface of oil bodies. The expression of oleosin-rhFGF9 was confirmed by polyacrylamide gel electrophoresis and western blotting. According to BCA and Enzyme-linked immunosorbent assay (ELISA) assay, the results show that the expression level of oleosin-rhFGF9 was 0.14% of oil body protein. The oil body bound oleosin-rhFGF9 showed mitogenic activity towards NIH3T3 cells in a methylthiazolyldiphenyl-tetrazolium bromide (MTT) assay. The efficacy of oil body bound oleosin-rhFGF9 in promoting hair growth and wound healing was investigated in C57BL/6 mice. In a hair regeneration experiment, 50 μg/μl oil body bound oleosin-rhFGF9 was applied to the dorsal skin of mice in the resting phase of the hair growth cycle. After 15 days, thicker hair and increased number of new hairs were seen compared with controls. Furthermore, the number of new hairs was greater compared with rhFGF9-treated mice. The hair follicles of mice treated with oil body bound oleosin-rhFGF9 expressed β-catenin more abundantly. In a wound healing experiment, dorsal skin wounds were topically treated with 50 μg/μl oil body bound oleosin-rhFGF9. Wound healing was quicker compared with mice treated with rhFGF9 and controls, especially in the earlier stages of healing.ConclusionsThe oil body bound oleosin-rhFGF9 promotes both hair growth and wound healing. It appears to promote hair growth, at least in part, by up-regulating β-catenin expression. The potential of oil body bound oleosin-rhFGF9 as an external drug can treat the alopecia and wounds or use in further clinical application.Electronic supplementary materialThe online version of this article (10.1186/s12896-018-0433-2) contains supplementary material, which is available to authorized users.

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Section: Resultsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Oil body bound oleosin-rhFGF9 fusion protein expressed in safflower (Carthamus tinctorius L.) stimulates hair growth and wound healing in mice

Cai

Wen

et al. 2018

BMC Biotechnol

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“…It could be a useful way of fortifying seeds with various proteins for health or to enhance seed protein yields. Yi et al ( 2015 ) used oleosin translational fusion technology (Van Rooijen and Moloney 1995a , b ) to produce human growth factor 9 fused to oleosin in Arabidopsis cells.…”

Section: Biotechnology—increasing Oil Productionmentioning

confidence: 99%

Oil body biogenesis and biotechnology in legume seeds

2017

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The seeds of many legume species including soybean, Pongamia pinnata and the model legume Medicago truncatula store considerable oil, apart from protein, in their cotyledons. However, as a group, legume storage strategies are quite variable and provide opportunities for better understanding of carbon partitioning into different storage products. Legumes with their ability to fix nitrogen can also increase the sustainability of agricultural systems. This review integrates the cell biology, biochemistry and molecular biology of oil body biogenesis before considering biotechnology strategies to enhance oil body biosynthesis. Cellular aspects of packaging triacylglycerol (TAG) into oil bodies are emphasized. Enhancing seed oil content has successfully focused on the up-regulation of the TAG biosynthesis pathways using overexpression of enzymes such as diacylglycerol acyltransferase1 and transcription factors such as WRINKLE1 and LEAFY COTYLEDON1. While these strategies are central, decreasing carbon flow into other storage products and maximizing the packaging of oil bodies into the cytoplasm are other strategies that need further examination. Overall there is much potential for integrating carbon partitioning, up-regulation of fatty acid and TAG synthesis and oil body packaging, for enhancing oil levels. In addition to the potential for integrated strategies to improving oil yields, the capacity to modify fatty acid composition and use of oil bodies as platforms for the production of recombinant proteins in seed of transgenic legumes provide other opportunities for legume biotechnology.

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“…The oil body could express exogenous proteins in oil crops seed, which can target exogenous proteins to the surface of oil bodies via covalent fusions with oleosin [ 16 ]. This has obvious advantages like lower production costs, more convenient storage, more stable heredity and higher output potential [ 17 – 19 ], compared with bacterial, fungal, insect and mammalian expression system.…”

Section: Introductionmentioning

confidence: 99%

A new nanoscale transdermal drug delivery system: oil body-linked oleosin-hEGF improves skin regeneration to accelerate wound healing

et al. 2018

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BackgroundEpidermal growth factor (EGF) can promote cell proliferation as well as migration, which is feasible in tissue wound healing. Oil bodies have been exploited as an important platform to produce exogenous proteins. The exogenous proteins were expressed in oil bodies from plant seeds. The process can reduce purification steps, thereby significantly reducing the purification cost. Mostly, the diameter of oil body particle ranges between 1.0 and 1.5 µm in the safflower seeds, however, it reduces to 700–1000 nm in the transgenic safflower seeds. The significant reduction of particle size in transgenic seeds is extremely beneficial to skin absorption.ResultsThe diameter of oil body in the transgenic safflower seeds was recorded in the range of 700–1000 nm. The smaller particle size improved their skin absorption. The expression level of oleosin-hEGF-hEGF in T3 transgenic seeds was highest at 69.32 mg/g of seeds. The oil body expressing oleosin-hEGF-hEGF had significant proliferative activity on NIH/3T3 cells and improved skin regeneration thereby accelerating wound healing in rats. The wound coverage rate exceeded 98% after treatment for 14 days with oil body expressing oleosin-hEGF-hEGF, while the saline without EGF group and wild type oil body group both showed less than 80%. The neonatal fibroblast and collagen were found to be increased in the safflower oil body expressing oleosin-hEGF-hEGF treatment group. TGF-β1, bFGF and VEGF were noted as important growth factors in the repair of cutaneous wounds. Their expression level increased after 4 and 7 day treatment, but decreased after 14 days. Therefore, it can promote skin regeneration to accelerate wounds healing.ConclusionsThe expression of oleosin-hEGF-hEGF in T3 transgenic seeds was 80.43 ng/μL oil body. It had significant proliferative activity on NIH/3T3 cells and improved skin regeneration to accelerate wound healing in rats. The expression process of TGF-β1, bFGF and VEGF increased at first and then gradually declined.Electronic supplementary materialThe online version of this article (10.1186/s12951-018-0387-5) contains supplementary material, which is available to authorized users.

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Expression of bioactive recombinant human fibroblast growth factor 9 in oil bodies of Arabidopsis thaliana

Cited by 18 publications

References 30 publications

Oil body bound oleosin-rhFGF9 fusion protein expressed in safflower (Carthamus tinctorius L.) stimulates hair growth and wound healing in mice

Oil body bound oleosin-rhFGF9 fusion protein expressed in safflower (Carthamus tinctorius L.) stimulates hair growth and wound healing in mice

Oil body biogenesis and biotechnology in legume seeds

A new nanoscale transdermal drug delivery system: oil body-linked oleosin-hEGF improves skin regeneration to accelerate wound healing

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