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Targeted gene deletion of miRNAs in mice by TALEN system

Academic Article
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Overview

authors

  • Takada, S.
  • Sato, T.
  • Ito, Y.
  • Yamashita, S.
  • Kato, T.
  • Kawasumi, M.
  • Kanai-Azuma, M.
  • Igarashi, A.
  • Kato, T.
  • Tamano, M.
  • Asahara, Hiroshi

publication date

  • October 2013

journal

  • PLoS One  Journal

abstract

  • Mice are among the most valuable model animal species with an enormous amount of heritage in genetic modification studies. However, targeting genes in mice is sometimes difficult, especially for small genes, such as microRNAs (miRNAs) and targeting genes in repeat sequences. Here we optimized the application of TALEN system for mice and successfully obtained gene targeting technique in mice for intergenic region and series of microRNAs. Microinjection of synthesized RNA of TALEN targeting each gene in one cell stage of embryo was carried out and injected oocytes were transferred into pseudopregnant ICR female mice, producing a high success rate of the targeted deletion of miRNA genes. In our condition, TALEN RNA without poly(A) tail worked better than that of with poly(A) tail. This mutated allele in miRNA was transmitted to the next generation, suggesting the successful germ line transmission of this targeting method. Consistent with our notion of miRNAs maturation mechanism, in homozygous mutant mice of miR-10a, the non- mutated strand of miRNAs expression was completely diminished. This method will lead us to expand and accelerate our genetic research using mice in a high throughput way.

subject areas

  • Animals
  • Base Sequence
  • DNA, Intergenic
  • DNA-Binding Proteins
  • Deoxyribonucleases, Type II Site-Specific
  • Embryo, Mammalian
  • Female
  • Gene Deletion
  • Gene Targeting
  • Genetic Engineering
  • Mice
  • Mice, Inbred ICR
  • MicroRNAs
  • Microinjections
  • Molecular Sequence Data
  • Oocytes
  • Recombinant Fusion Proteins
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Identity

PubMed Central ID

  • PMC3797721

International Standard Serial Number (ISSN)

  • 1932-6203

Digital Object Identifier (DOI)

  • 10.1371/journal.pone.0076004

PubMed ID

  • 24146809
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Additional Document Info

start page

  • e76004

volume

  • 8

issue

  • 10

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