Establishment of a transgenic mouse model of corneal dystrophy overexpressing human BIGH3

Liao,Xin; Cui,Hongping; Wang,Fang

doi:10.3892/ijmm.2013.1480

Establishment of a transgenic mouse model of corneal dystrophy overexpressing human BIGH3

Authors:
- Xin Liao
- Hongping Cui
- Fang Wang
View Affiliations

Affiliations: Department of Ophthalmology, Shanghai Tenth People's Hospital Affiliated to Tongji University, Shanghai 200072, P.R. China, Department of Ophthalmology, Shanghai East Hospital Affiliated to Tongji University School of Medicine, Shanghai 200120, P.R. China
Published online on: September 4, 2013 https://doi.org/10.3892/ijmm.2013.1480
Pages: 1110-1114

Metrics: Total Views: 0 (Spandidos Publications: | PMC Statistics: )

Metrics: Total PDF Downloads: 0 (Spandidos Publications: | PMC Statistics: )

Cited By (CrossRef): 0 citations Loading Articles...

Abstract

This study aimed to establish a transgenic mouse model of corneal dystrophy (CD) overexpressing the human transforming growth factor, β-induced, 68 kDa (TGFBI, also known as BIGH3) gene. A purified and linearized recombinant plasmid carrying the expression cassette BIGH3‑IRES‑EGFP was microinjected into the pronuclei of C57BL/6J mouse fertilized eggs under the control of the phosphoglycerate kinase (PGK) promoter. The expression of human BIGH3 in the transgenic mice was confirmed by PCR using DNA extracted from tail tissue. Four founder transgenic mice were identified by PCR and the increased expression of BIGH3 was observed in the corneas of the transgenic mice by RT‑PCR and western blot analysis. The abnormal corneas with central opacity were observed in the transgenic mice by corneal photography. We concluded that the exogenous gene, BIGH3, was integrated successfully into the mouse genome through microinjection. In addition, the phenotype observed in this BIGH3 transgenic mouse model was similar to CD. Therefore, this transgenic model may prove useful in the investigation of the pathogenesis of CD.

View Figures

View References

1	Eiberg H, Moller HU, Berendt I and Mohr J: Assignment of granular corneal dystrophy Groenouw type I (CDGG1) to chromosome 5q. Eur J Hum Genet. 2:132–138. 1994.PubMed/NCBI
2	Yu P, Gu Y, Yang Y, et al: A clinical and molecular-genetic analysis of Chinese patients with lattice corneal dystrophy and novel Thr538Pro mutation in the TGFBI (BIGH3) gene. J Genet. 85:73–76. 2006. View Article : Google Scholar : PubMed/NCBI
3	Paliwal P, Sharma A, Tandon R, et al: TGFBI mutation screening and genotype-phenotype correlation in north Indian patients with corneal dystrophies. Mol Vis. 16:1429–1438. 2010.PubMed/NCBI
4	Vincent AL, de Karolyi B, Patel DV, et al: TGFBI mutational analysis in a New Zealand population of inherited corneal dystrophy patients. Br J Ophthalmol. 94:836–842. 2010. View Article : Google Scholar : PubMed/NCBI
5	Korvatska E, Munier FL, Djemai A, et al: Mutation hot spots in 5q31-linked corneal dystrophies. Am J Hum Genet. 62:320–324. 1998. View Article : Google Scholar : PubMed/NCBI
6	Skonier J, Bennett K, Rothwell V, et al: Beta-igh3: a transforming growth factor-beta responsive gene encoding a secreted protein that inhibits cell attachment in vitro and suppresses the growth of CHO cells in nude mice. DNA Cell Biol. 13:571–584. 1994. View Article : Google Scholar : PubMed/NCBI
7	Grunauer-Kloevekorn C, Braeutigam S, Weidle E, et al: Molecular genetic and histopathological examinations for genotype-phenotype analysis in patients with TGFBI-linked corneal dystrophy. Klin Monbl Augenheilkd. 223:829–836. 2006.(In German).
8	Kannabiran C and Klintworth GK: TGFBI gene mutations in corneal dystrophies. Hum Mutat. 27:615–625. 2006. View Article : Google Scholar : PubMed/NCBI
9	Patel DA, Chang SH, Harocopos GJ, et al: Granular and lattice deposits in corneal dystrophy caused by R124C mutation of TGFBIp. Cornea. 11:1215–1222. 2010. View Article : Google Scholar : PubMed/NCBI
10	Gruenauer-Kloevekorn C, Clausen I, Weidle E, et al: TGFBI (BIGH3) gene mutations in German families: two novel mutations associated with unique clinical and histopathological findings. Br J Ophthalmol. 93:932–937. 2009. View Article : Google Scholar : PubMed/NCBI
11	Grunauer-Kloevekorn C, Brautigam S, Wolter-Roessler M, et al: Molecular genetic analysis of the BIGH3 gene in lattice type I (Biber-Haab-Dimmer) and granular type II (Avellino) corneal dystrophy: is indirect mutation analysis for hot spots recommended? Klin Monbl Augenheilkd. 222:1017–1023. 2005.(In German).
12	Munier FL, Korvatska E, Djemai A, et al: Kerato-epithelin mutations in four 5q31- linked corneal dystrophies. Nat Genet. 15:247–251. 1997. View Article : Google Scholar : PubMed/NCBI
13	Kim JE, Han MS, Bae YC, et al: Anterior segment dysgenesis after overexpression of transforming growth factor-beta-induced gene, beta igh3, in the mouse eye. Mol Vis. 13:1942–1952. 2007.PubMed/NCBI
14	Bustamante M, Tasinato A, Maurer F, et al: Overexpression of a mutant form of TGFBI/BIGH3 induces retinal degeneration in transgenic mice. Mol Vis. 14:1129–1137. 2008.PubMed/NCBI
15	Pey AL, Mesa-Torres N, Chiarelli LR and Valentini G: Structural and energetic basis of protein kinetic destabilization in human phosphoglycerate kinase 1 deficiency. Biochemistry. 52:1160–1170. 2013. View Article : Google Scholar : PubMed/NCBI
16	Schay G, Herenyi L, Fidy J and Osvath S: Role of domain interactions in the collective motion of phosphoglycerate kinase. Biophys J. 104:677–682. 2013. View Article : Google Scholar : PubMed/NCBI
17	Lei Y and Liu YG: Isolation, sequence identification and tissue expression profile of a novel sheep gene-PGK1. Res J Biotechnol. 8:38–41. 2013.
18	Singer-Sam J, Keith DH, Tani K, et al: Sequence of the promoter region of the gene for human X-linked 3-phosphoglycerate kinase. Gene. 32:409–417. 1984. View Article : Google Scholar : PubMed/NCBI
19	Takada T, Lida K, Awaji T, et al: Selective production of transgenic mice using green fluorescent protein as a marker. Nat Biotechnol. 15:458–461. 1997. View Article : Google Scholar : PubMed/NCBI
20	Mizuguchi H, Xu Z, Ishii-Watabe A, et al: IRES-dependent second gene expression is significantly lower than cap-dependent first gene expression in a bicistronic vector. Mol Ther. 1:376–382. 2000. View Article : Google Scholar : PubMed/NCBI
21	Borman AM, Bailly JL, Girard M and Kean KM: Picornavirus internal ribosome entry segments: comparison of translation efficiency and the requirements for optimal internal initiation of translation in vitro. Nucleic Acids Res. 23:3656–3663. 1995. View Article : Google Scholar
22	Cao HQ and Ding JF: Construction strategy of polygene co-expression vectors. Foreign Med Sci (Molecular Biology Section). 24:1–4. 2000.