Transgenic Research

, Volume 20, Issue 5, pp 975–988

High efficiency of BRCA1 knockout using rAAV-mediated gene targeting: developing a pig model for breast cancer

  • Yonglun Luo
  • Juan Li
  • Ying Liu
  • Lin Lin
  • Yutao Du
  • Shengting Li
  • Huanming Yang
  • Gábor Vajta
  • Henrik Callesen
  • Lars Bolund
  • Charlotte Brandt Sørensen
Original Paper

DOI: 10.1007/s11248-010-9472-8

Cite this article as:
Luo, Y., Li, J., Liu, Y. et al. Transgenic Res (2011) 20: 975. doi:10.1007/s11248-010-9472-8
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Abstract

Germline inactivating mutations of the breast cancer associated gene 1 (BRCA1) predispose to breast cancer and account for most cases of familiar breast and/or ovarian cancer. The pig is an excellent model for medical research as well as testing of new methods and drugs for disease prevention and treatment. We have generated cloned BRCA1 knockout (KO) Yucatan miniature piglets by targeting exon 11 using recombinant adeno-associated virus (rAAV)-mediated gene targeting and somatic cell nuclear transfer by Handmade Cloning (HMC). We found a very high targeting rate of rAAV-mediated BRCA1 KO. Approximately 35% of the selected cells were BRCA1 targeted. One BRCA1 KO cell clone (5D1), identified by PCR and Southern blot, was used as nuclear donor for HMC. Reconstructed embryos were transferred to three recipient sows which gave birth to 8 piglets in total. Genotyping identified seven piglets as BRCA1 heterozygotes (BRCA1+/∆11), and one as wild type. The BRCA1 expression was decreased at the mRNA level in BRCA1+/∆11 fibroblasts. However, all BRCA1+/∆11 piglets died within 18 days after birth. The causes of perinatal mortality remain unclear. Possible explanations may include a combination of the BRCA1 haploinsufficiency, problems of epigenetic reprogramming, presence of the marker gene, single cell clone effects, and/or the special genetic background of the minipigs.

Keywords

BRCA1 Breast cancer rAAV Gene targeting Minipig 

Supplementary material

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Supplementary material 4 (TIFF 1135 kb)

Copyright information

© Springer Science+Business Media B.V. 2010

Authors and Affiliations

  • Yonglun Luo
    • 1
    • 3
  • Juan Li
    • 2
  • Ying Liu
    • 2
  • Lin Lin
    • 2
    • 3
  • Yutao Du
    • 3
  • Shengting Li
    • 4
  • Huanming Yang
    • 3
  • Gábor Vajta
    • 2
  • Henrik Callesen
    • 2
  • Lars Bolund
    • 1
    • 3
  • Charlotte Brandt Sørensen
    • 1
  1. 1.Department of Human GeneticsAarhus UniversityAarhus CDenmark
  2. 2.Section of Population Genetics and Embryology, Department of Genetics and Biotechnology, Research Centre FoulumAarhus UniversityTjeleDenmark
  3. 3.BGI Shenzhen (HuaDa Shenzhen)ShenzhenChina
  4. 4.Bioinformatics Research CentreAarhus UniversityAarhus CDenmark

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