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

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.

[1]  L. Chiarelli,et al.  Structural and energetic basis of protein kinetic destabilization in human phosphoglycerate kinase 1 deficiency. , 2013, Biochemistry.

[2]  S. Osváth,et al.  Role of domain interactions in the collective motion of phosphoglycerate kinase. , 2013, Biophysical journal.

[3]  L. Yonggang,et al.  Isolation, Sequence Identification and Tissue Expressio Profile of a Novel Sheep Gene-SERPINF1 , 2011 .

[4]  G. Harocopos,et al.  Granular and Lattice Deposits in Corneal Dystrophy Caused by R124C Mutation of TGFBIp , 2010, Cornea.

[5]  P. Kaur,et al.  TGFBI mutation screening and genotype-phenotype correlation in north Indian patients with corneal dystrophies , 2010, Molecular vision.

[6]  D. Patel,et al.  TGFBI mutational analysis in a New Zealand population of inherited corneal dystrophy patients , 2009, British Journal of Ophthalmology.

[7]  F. Tost,et al.  TGFBI (BIGH3) gene mutations in German families: two novel mutations associated with unique clinical and histopathological findings , 2008, British Journal of Ophthalmology.

[8]  Y. Arsenijévic,et al.  Overexpression of a mutant form of TGFBI/BIGH3 induces retinal degeneration in transgenic mice , 2008, Molecular vision.

[9]  In‐San Kim,et al.  Anterior segment dysgenesis after overexpression of transforming growth factor-beta-induced gene, beta igh3, in the mouse eye. , 2007 .

[10]  P. Yu,et al.  A clinical and molecular-genetic analysis of Chinese patients with lattice corneal dystrophy and novel Thr538Pro mutation in theTGFBI (BIGH3) gene , 2006, Journal of Genetics.

[11]  C. Grünauer-Kloevekorn,et al.  Molekulargenetische und histopathologische Untersuchungen zur Genotyp-Phänotyp-Analyse bei Patienten mit TGFBI-gekoppelten Hornhautdystrophien , 2006 .

[12]  C. Kannabiran,et al.  TGFBI gene mutations in corneal dystrophies , 2006, Human mutation.

[13]  P. Yu,et al.  A clinical and molecular-genetic analysis of Chinese patients with lattice corneal dystrophy and novel Thr538Pro mutation in the TGFBI (BIGH3) gene. , 2006, Journal of genetics.

[14]  F. Tost,et al.  [Molecular genetic and histopathological examinations for genotype-phenotype analysis in patients with TGFBI-linked corneal dystrophy]. , 2006, Klinische Monatsblatter fur Augenheilkunde.

[15]  C. Grünauer-Kloevekorn,et al.  Molekulargenetische Analyse des BIGH3-Gens bei gittriger Hornhautdystrophie Typ I (Biber-Haab-Dimmer) und bei bröckliger Hornhautdystrophie Typ II (Avellino): Erlauben Hot Spots einen indirekten Mutationsnachweis? , 2005 .

[16]  F. Tost,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?]. , 2005, Klinische Monatsblatter fur Augenheilkunde.

[17]  H. Mizuguchi,et al.  IRES-dependent second gene expression is significantly lower than cap-dependent first gene expression in a bicistronic vector. , 2000, Molecular therapy : the journal of the American Society of Gene Therapy.

[18]  W. Culbertson,et al.  Mutation hot spots in 5q31-linked corneal dystrophies. , 1998, American journal of human genetics.

[19]  D. Schorderet,et al.  Kerato-epithelin mutations in four 5q31-linked corneal dystrophies. , 1997 .

[20]  Sumio Sugano,et al.  Selective production of transgenic mice using green fluorescent protein as a marker , 1997, Nature Biotechnology.

[21]  D. Paslier,et al.  Kerato-epithelin mutations in four 5q31-linked corneal dystrophies , 1997, Nature Genetics.

[22]  K. Kean,et al.  Picornavirus internal ribosome entry segments: comparison of translation efficiency and the requirements for optimal internal initiation of translation in vitro. , 1995, Nucleic acids research.

[23]  Hans Marquardt,et al.  βig-h3: A Transforming Growth Factor-β-Responsive Gene Encoding a Secreted Protein That Inhibits Cell Attachment In Vitro and Suppresses the Growth of CHO Cells in Nude Mice , 1994 .

[24]  C. Disteche,et al.  beta ig-h3: 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. , 1994, DNA and cell biology.

[25]  H. Eiberg,et al.  Assignment of Granular Corneal Dystrophy Groenouw Type I (CDGG1) to Chromosome 5q , 1994, European journal of human genetics : EJHG.

[26]  A. Riggs,et al.  Sequence of the promoter region of the gene for human X-linked 3-phosphoglycerate kinase. , 1984, Gene.