Combinational Treatment of Trichostatin A and Vitamin C Improves the Efficiency of Cloning Mice by Somatic Cell Nuclear Transfer

J Vis Exp. 2018 Apr 26:(134):57036. doi: 10.3791/57036.

Abstract

Somatic cell nuclear transfer (SCNT) provides a unique opportunity to directly produce a cloned animal from a donor cell, and it requires the use of skillful techniques. Additionally, the efficiencies of cloning have remained low since the successful production of cloned animals, especially mice. There have been many attempts to improve the cloning efficiency, and trichostatin A (TSA), a histone deacetylase inhibitor, has been widely used to enhance the efficiency of cloning. Here, we report a dramatically improved cloning method in mice. This somatic cell nuclear transfer method involves usage of Hemagglutinating virus of Japan Envelope (HVJ-E), which enables easy manipulation. Moreover, the treatment using two small molecules, TSA and vitamin C (VC), with deionized bovine serum albumin (dBSA), is highly effective for embryonic development. This approach requires neither additional injection nor genetic manipulation, and thus presents a simple, suitable method for practical use. This method could become a technically feasible approach for researchers to produce genetically modified animals from cultured cells. Furthermore, it might be a useful way for the rescue of endangered animals via cloning.

Publication types

  • Research Support, Non-U.S. Gov't
  • Video-Audio Media

MeSH terms

  • Animals
  • Ascorbic Acid / pharmacology
  • Ascorbic Acid / therapeutic use*
  • Cloning, Organism / methods*
  • Embryonic Development / drug effects
  • Female
  • Hydroxamic Acids / pharmacology
  • Hydroxamic Acids / therapeutic use*
  • Male
  • Mice
  • Nuclear Transfer Techniques / trends*
  • Pregnancy

Substances

  • Hydroxamic Acids
  • trichostatin A
  • Ascorbic Acid