Identification of genes important for cutaneous function revealed by a large scale reverse genetic screen in the mouse.

Tia Ditommaso,Karen P Steel,Fiona M Watt,Denny L Cottle,Ramiro Ramirez-Solis,Lynelle K Jones,Valerie E Vancollie,Anna-Karin Gerdin,Ian M Smyth,Jacqueline K White,Allan Bradley,John P Sundberg,Gregory S Barsh

doi:10.1371/journal.pgen.1004705

Tia Ditommaso, Karen P Steel + Show 11 more

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https://doi.org/10.1371/journal.pgen.1004705

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Abstract

The skin is a highly regenerative organ which plays critical roles in protecting the body and sensing its environment. Consequently, morbidity and mortality associated with skin defects represent a significant health issue. To identify genes important in skin development and homeostasis, we have applied a high throughput, multi-parameter phenotype screen to the conditional targeted mutant mice generated by the Wellcome Trust Sanger Institute's Mouse Genetics Project (Sanger-MGP). A total of 562 different mouse lines were subjected to a variety of tests assessing cutaneous expression, macroscopic clinical disease, histological change, hair follicle cycling, and aberrant marker expression. Cutaneous lesions were associated with mutations in 23 different genes. Many of these were not previously associated with skin disease in the organ (Mysm1, Vangl1, Trpc4ap, Nom1, Sparc, Farp2, and Prkab1), while others were ascribed new cutaneous functions on the basis of the screening approach (Krt76, Lrig1, Myo5a, Nsun2, and Nf1). The integration of these skin specific screening protocols into the Sanger-MGP primary phenotyping pipelines marks the largest reported reverse genetic screen undertaken in any organ and defines approaches to maximise the productivity of future projects of this nature, while flagging genes for further characterisation.

Highlights

Recent advances in the high throughput production of targeted mutant mice using gene targeting in ES cells have made it possible to rapidly and effectively produce knockout mouse lines for use in phenotypic screening [1,2,3,4]
Central to recent community efforts has been the Wellcome Trust Sanger Institute’s Mouse Genetics Project (Sanger-MGP), which is undertaking a high throughput reverse genetic screen using the knockout ES cells generated by the International Knockout Mouse Consortium, which has merged with the International Phenotyping Mouse Consortium
The immediate connection of a phenotype to a mutated gene represents a paradigm shift in our ability to explore gene function. This study utilized such a screening approach to investigate the genetic contribution to skin development and homeostasis

Summary

Introduction

Recent advances in the high throughput production of targeted mutant mice using gene targeting in ES cells have made it possible to rapidly and effectively produce knockout mouse lines for use in phenotypic screening [1,2,3,4]. Such approaches are beginning to yield significant insights into mammalian gene function [1,5].

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