Targeted disruption of Tbc1d20 with zinc-finger nucleases causes cataracts and testicular abnormalities in mice

BMC Genetics

Duska J Sidjanin¹  Aron Geurts¹  Allison D Ebert²  Adam Ronchetti²  Ryan P Liegel²  Anna Kyunglim Park² 

[1] Human and Molecular Genetics Center, Medical College of Wisconsin, 8701 Watertown Plank, Milwaukee 53226, WI, USA;Department of Cell Biology, Neurobiology and Anatomy, Medical College of Wisconsin, 8701 Watertown Plank, Milwaukee 53226, WI, USA

关键词: Warburg Micro Syndrome;    Spermatogenesis;    Blind-sterile;    Zinc-finger nuclease;    Loss-of-function;    TBC1D20;   

Others  :  1085132
DOI  :  10.1186/s12863-014-0135-2

 received in 2014-10-09, accepted in 2014-11-24,  发布年份 2014

【 摘 要 】

Background

Loss-of-function mutations in TBC1D20 cause Warburg Micro syndrome 4 (WARBM4), which is an autosomal recessive syndromic disorder characterized by eye, brain, and genital abnormalities. Blind sterile (bs) mice carry a Tbc1d20-null mutation and exhibit cataracts and testicular phenotypes similar to those observed in WARBM4 patients. In addition to TBC1D20, mutations in RAB3GAP1, RAB3GAP2 and RAB18 cause WARBM1-3 respectively. However, regardless of which gene harbors the causative mutation, all individuals affected with WARBM exhibit indistinguishable clinical presentations. In contrast, bs, Rab3gap1^-/-, and Rab18^-/- mice exhibit distinct phenotypes; this phenotypic variability of WARBM mice was previously attributed to potential compensatory mechanisms. Rab3gap1^-/- and Rab18^-/- mice were genetically engineered using standard approaches, whereas the Tbc1d20 mutation in the bs mice arose spontaneously. There is the possibility that another unidentified mutation within the bs linkage disequilibrium may be contributing to the bs phenotypes and thus contributing to the phenotypic variability in WARBM mice. The goal of this study was to establish the phenotypic consequences in mice caused by the disruption of the Tbc1d20 gene.

Results

The zinc finger nuclease (ZFN) mediated genomic editing generated a Tbc1d20 c.[418_426del] deletion encoding a putative TBC1D20-ZFN protein with an in-frame p.[H140_Y143del] deletion within the highly conserved TBC domain. The evaluation of Tbc1d20^ZFN/ZFN eyes identified severe cataracts and thickened pupillary sphincter muscle. Tbc1d20^ZFN/ZFN males are infertile and the analysis of the seminiferous tubules identified disrupted acrosomal development. The compound heterozygote Tbc1d20^ZFN/bs mice, generated from an allelic bs/+ X Tbc1d20^ZFN/+ cross, exhibited cataracts and aberrant acrosomal development indicating a failure to complement.

Conclusions

Our findings show that the disruption of Tbc1d20 in mice results in cataracts and aberrant acrosomal formation, thus establishing bs and Tbc1d20^ZFN/ZFN as allelic variants. Although the WARBM molecular disease etiology remains unclear, both the bs and Tbc1d20^ZFN/ZFN mice are excellent model organisms for future studies to establish TBC1D20-mediated molecular and cellular functions.

【 授权许可】

   
2014 Park et al.; licensee BioMed Central Ltd.

【 图 表 】

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