September 2016
Volume 57, Issue 12
Open Access
ARVO Annual Meeting Abstract  |   September 2016
Knock-down of Tropomyosin 2 created by CRISPR-CAS9 in mouse induces age-related-cataract
Author Affiliations & Notes
  • Eri Kubo
    Department of Ophthalmology, Kanazawa Medical University, Ishikawa, Japan
  • Teppei Shibata
    Department of Ophthalmology, Kanazawa Medical University, Ishikawa, Japan
  • Shinsuke Shibata
    Department of Ophthalmology, Kanazawa Medical University, Ishikawa, Japan
  • Hiroshi Sasaki
    Department of Ophthalmology, Kanazawa Medical University, Ishikawa, Japan
  • Etsuko Kiyokawa
    Department of Oncogenic Pathology, Kanazawa Medical University, Ishikawa, Japan
  • Masato Ikawa
    Research Institute for Microbial Diseases, Osaka University, Osaka, Japan
  • Dhirendra P Singh
    Department of Ophthalmology and Visual Sciences, University of Nebraska Medical Center, Omaha, Nebraska, United States
  • Footnotes
    Commercial Relationships   Eri Kubo, Ono Pharmaceutical Co Ltd (F); Teppei Shibata, Ono Pharmaceutical Co Ltd (F); Shinsuke Shibata, None; Hiroshi Sasaki, Ono Pharmaceutical Co Ltd (F); Etsuko Kiyokawa, None; Masato Ikawa, None; Dhirendra Singh, None
  • Footnotes
    Support  JSPS Grants-in-Aid for Scientific Research C 23592588
Investigative Ophthalmology & Visual Science September 2016, Vol.57, No Pagination Specified. doi:
  • Views
  • Share
  • Tools
    • Alerts
      ×
      This feature is available to authenticated users only.
      Sign In or Create an Account ×
    • Get Citation

      Eri Kubo, Teppei Shibata, Shinsuke Shibata, Hiroshi Sasaki, Etsuko Kiyokawa, Masato Ikawa, Dhirendra P Singh; Knock-down of Tropomyosin 2 created by CRISPR-CAS9 in mouse induces age-related-cataract. Invest. Ophthalmol. Vis. Sci. 201657(12):.

      Download citation file:


      © ARVO (1962-2015); The Authors (2016-present)

      ×
  • Supplements
Abstract

Purpose : The tropomyosin (Tpm) family of cytoskeleton proteins is involved in regulating and stabilizing actin microfilaments (F-actin). Previously we demonstrated that elevated expression of Tpm1α/2β was related to progression of rat posterior capsule opacification (PCO) after cataract surgery. In this study, we investigated the role of Tpm2 in lens by generating Tpm2 hetero-knock-out (Tpm2+/-) mice, using the clustered regularly interspaced short palindromic repeat/Cas9 (CRISPR/Cas9) system.

Methods : Experiments accorded with National Institutes of Health Guide for the Care and Use of Laboratory Animals. We generated CRISPR/CAS9 -mediated Tpm2 mutations in germline C57BL/6NCr mice. In mice aged 7-, 16- and 26-weeks old, lenses were extracted (n=6eyes), and observed by microscope. The expression levels of mRNA and protein were examined by western blot and real-time RT PCR, respectively. Tissues in paraffin sections were histologically analyzed.

Results : Tpm2+/- mice were selected because the Tpm2 gene null-knockout mouse exhibits viviparous lethality. At 7- and 16-weeks old, lenses were transparent in both Tpm2+/- and wild type (WT) mice. However, at 26-weeks old cataract was observed in the posterior part of the lens or annular opacity in Tpm2+/- mice, not in WT mice.

Conclusions : Tpm2+/- mice can be created using the CRISPR/Cas9 system. Tpm2 is involved in the progression of age-related cataract. The present results indicate that low levels of Tpm2 in the lens contribute the age-related cataract development. This is a useful model to study cataractogenesis and PCO.

This is an abstract that was submitted for the 2016 ARVO Annual Meeting, held in Seattle, Wash., May 1-5, 2016.

×
×

This PDF is available to Subscribers Only

Sign in or purchase a subscription to access this content. ×

You must be signed into an individual account to use this feature.

×