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In-vitro Mutagenesis01:16

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To learn more about the function of a gene, researchers can observe what happens when the gene is inactivated or “knocked out,” by creating genetically engineered knockout animals. Knockout mice have been particularly useful as models for human diseases such as cancer, Parkinson’s disease, and diabetes.
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Related Experiment Video

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Production of Apolipoprotein C-III Knockout Rabbits using Zinc Finger Nucleases
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Generation of RUNX3 knockout pigs using CRISPR/Cas9-mediated gene targeting.

J-T Kang1, J Ryu2, B Cho1

  • 1MGENPLUS Biotechnology Research Institute, Seoul, Korea.

Reproduction in Domestic Animals = Zuchthygiene
|October 4, 2016
PubMed
Summary

Researchers created genetically engineered pigs lacking the RUNX3 tumor suppressor gene using CRISPR/Cas9 technology. These RUNX3 knockout pigs serve as a valuable model for studying human gastric cancer and developing new therapies.

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Area of Science:

  • Genetics and Genomics
  • Animal Models
  • Cancer Research

Background:

  • Pigs offer anatomical and physiological similarities to humans, making them ideal for cancer research.
  • The development of genetically engineered pigs has been a bottleneck for recapitulating human cancer progression.
  • Genome editing technologies like CRISPR/Cas9 enable efficient generation of genetically modified pigs.

Purpose of the Study:

  • To generate a pig model for human gastric cancer research using CRISPR/Cas9.
  • To target the RUNX3 tumor suppressor gene for creating knockout pigs.
  • To assess the utility of RUNX3 knockout pigs in cancer research and therapy development.

Main Methods:

  • Generated RUNX3 knockout cell lines (monoallelic or biallelic) in foetal fibroblast cells using CRISPR/Cas9.
  • Utilized somatic cell nuclear transfer with genetically modified cells for embryo creation.
  • Embryo transfer into surrogates to obtain live piglets.

Main Results:

  • Successfully produced four live RUNX3 knockout piglets.
  • Confirmed the absence of RUNX3 protein in the internal organs of the knockout piglets.
  • Demonstrated the efficacy of CRISPR/Cas9 in targeted genomic mutation induction.

Conclusions:

  • CRISPR/Cas9 technology is effective for creating specific gene knockouts in pigs.
  • RUNX3 knockout pigs represent a valuable resource for human gastric cancer research.
  • These pigs can aid in the development of innovative cancer treatments.