title: Expression of the HSF4 DNA binding domain-EGFP hybrid gene recreates early childhood lamellar cataract in transgenic mice.
abstract: OBJECTIVE
  The clinical management of cataracts in infancy involves surgical removal of the lens to ensure transmission of light to the retina, which is essential for normal neural development of the infant. This surgery, however, entails a lifelong follow-up and impaired vision. To our knowledge, no animal models recapitulate human lamellar opacities, the most prevalent form of early childhood cataracts. We present data on the recreation of the human lamellar cataract phenotype in transgenic mice.
  
  
  METHODS
  Mutations in the DNA binding domain (DBD) of the heat shock transcription factor 4 (HSF4) are known to be associated with early childhood autosomal dominant lamellar cataract. We used bacterial artificial chromosome (BAC) transgenesis to express a hybrid gene: Hsf4 (DBD)-enhanced green fluorescent protein (EGFP), by recombineering EGFP sequences into the DBD of the Hsf4 gene, to interfere with the DNA binding properties of Hsf4.
  
  
  RESULTS
  We recapitulated the human lamellar cataract, in its temporal as well as spatial presentation, within the transgenic mouse lens. This phenotype was reproduced faithfully using four different BACs, indicating that EGFP can be used to target transcription factor function in transgenic mice. Molecular and cell biological examination of early postnatal transgenic lens reveals impairment of secondary fiber cell differentiation.
  
  
  CONCLUSIONS
  Recreation of the human lamellar cataract phenotype in mice allows investigation of this human pathology at a level not possible previously and points to the relevance of fiber cell heterogeneity dictated by fiber cell-specific gene activity in the biogenesis of the lamellar cataract.
authors.affiliation: Jules Stein Eye Institute, Geffen School of Medicine at University of California Los Angeles, Los Angeles, California, United States.
authors.forename: Rajendra K
authors.initials: RK
authors.lastname: Gangalum
authors.affiliation: Jules Stein Eye Institute, Geffen School of Medicine at University of California Los Angeles, Los Angeles, California, United States.
authors.forename: Zhe
authors.initials: Z
authors.lastname: Jing
authors.affiliation: Jules Stein Eye Institute, Geffen School of Medicine at University of California Los Angeles, Los Angeles, California, United States.
authors.forename: Ankur M
authors.initials: AM
authors.lastname: Bhat
authors.affiliation: Jules Stein Eye Institute, Geffen School of Medicine at University of California Los Angeles, Los Angeles, California, United States Department of Molecular and Medical Pharmacology, Geffen School of Medicine at University of California Los Angeles, Los Angeles, California, United States Molecular Biology Institute and Brain Research Institute, University of California Los Angeles, Los Angeles, California, United States.
authors.forename: Josh
authors.initials: J
authors.lastname: Lee
authors.affiliation: Department of Molecular and Medical Pharmacology, Geffen School of Medicine at University of California Los Angeles, Los Angeles, California, United States.
authors.forename: Yoshiko
authors.initials: Y
authors.lastname: Nagaoka
authors.affiliation: Molecular Biology Institute and Brain Research Institute, University of California Los Angeles, Los Angeles, California, United States.
authors.forename: Sophie X
authors.initials: SX
authors.lastname: Deng
authors.affiliation: Department of Molecular and Medical Pharmacology, Geffen School of Medicine at University of California Los Angeles, Los Angeles, California, United States.
authors.forename: Meisheng
authors.initials: M
authors.lastname: Jiang
authors.affiliation: Jules Stein Eye Institute, Geffen School of Medicine at University of California Los Angeles, Los Angeles, California, United States Molecular Biology Institute and Brain Research Institute, University of California Los Angeles, Los Angeles, California, United States.
authors.forename: Suraj P
authors.initials: SP
authors.lastname: Bhat
mesh_terms: D000818:Animals; D002386:Cataract / Q000235:genetics* / Q000378:metabolism / Q000473:pathology; D002478:Cells, Cultured; D002648:Child; D004247:DNA / Q000235:genetics*; D004268:DNA-Binding Proteins / Q000096:biosynthesis / Q000235:genetics*; D004195:Disease Models, Animal; D018507:Gene Expression Regulation, Developmental; D018773:Genes, erbB-1 / Q000235:genetics*; D005838:Genotype; D000076249:Heat Shock Transcription Factors; D006801:Humans; D015151:Immunoblotting; D051379:Mice; D008822:Mice, Transgenic; D060888:Real-Time Polymerase Chain Reaction; D014157:Transcription Factors / Q000096:biosynthesis / Q000235:genetics*
chemical_list: D004268:DNA-Binding Proteins; D000076249:Heat Shock Transcription Factors; C484462:Hsf4 protein, mouse; D014157:Transcription Factors; D004247:DNA
keywords: BACs; HSF4; cataract; lens; transgenic mice