Transgenic Research

, Volume 22, Issue 2, pp 417–423

Efficient production of transgenic chickens based on piggyBac

  • Xiaojuan Liu
  • Ning Li
  • Xiaoxiang Hu
  • Ran Zhang
  • Qingyuan Li
  • Dainan Cao
  • Tongxin Liu
  • Yaqiong Zhang
  • Xiaofang Liu
Brief Communication

DOI: 10.1007/s11248-012-9642-y

Cite this article as:
Liu, X., Li, N., Hu, X. et al. Transgenic Res (2013) 22: 417. doi:10.1007/s11248-012-9642-y

Abstract

Transgenic techniques in chickens have been developed much more slowly than in mammals due to chickens’ unique reproduction mechanism. Retroviral methods have been the most successful. piggyBac (PB) is a transposon that has a 13 bp perfect terminal invert repeat sequence. PB can be inserted into TTAA sites and can also be precisely excised in mammals. Therefore, we have selected PB as a candidate to establish a new method to produce transgenic chickens. We constructed three donor vectors (ZGl-neo, ZGm-neo and ZGs-neo) expressing a GFP marker-gene and a neomycin resistant gene based on PB. We co-transfected each donor vector with a helper vector (CAG-PBase). We found that ZGl-neo was the most efficient PB vector. This vector could insert into TTAA sites in DF-1 cells. PB vectors were microinjected into sub-germinal cavity of newly laid eggs, and electroporation was then performed with a 20-V pulse for 5 cycles of 50 ms on and 100 ms off. GFP was expressed in different tissues of the embryos, including the gonads. Twenty-two chickens hatched after microinjection with compounds ZGl-neo and CAG-PBase (3:1). When we screened the blood DNA, 73 % (16/22) of the individuals were positive. Thirteen of the chickens grew to adulthood, 11 of which were males. 40 % (4/10) of the individuals were semen positive, and their copy numbers ranged from 0.05 to 0.21 (0.11, 0.21, 0.05, 0.06). No G1 offspring containing the integrated transposon were produced. We conclude that the PB transposon system is a novel useful tool for the efficient production of transgenic chickens.

Keywords

PiggyBac Transgenic chicken Electroporation Microinjection 

Supplementary material

11248_2012_9642_MOESM1_ESM.docx (17 kb)
Supplementary material 1 (DOCX 17 kb)
11248_2012_9642_MOESM2_ESM.tif (69 kb)
Supplement Fig. 1 The distribution of PB’s target sites on the chromosome. The horizontal axis represented the chromosome, the vertical Axis represented the clone number. (TIFF 68 kb)
11248_2012_9642_MOESM3_ESM.tif (1.5 mb)
Supplement Fig. 2 GFP was expressed in the testis of adult cock. A, control cock; B, transgenic cock. (TIFF 1499 kb)
11248_2012_9642_MOESM4_ESM.xlsx (14 kb)
Supplementary material 4 (XLSX 13 kb)
11248_2012_9642_MOESM5_ESM.xlsx (12 kb)
Supplementary material 5 (XLSX 12 kb)
11248_2012_9642_MOESM6_ESM.xlsx (14 kb)
Supplementary material 6 (XLSX 14 kb)

Copyright information

© Springer Science+Business Media B.V. 2012

Authors and Affiliations

  • Xiaojuan Liu
    • 1
  • Ning Li
    • 1
  • Xiaoxiang Hu
    • 1
  • Ran Zhang
    • 1
  • Qingyuan Li
    • 1
  • Dainan Cao
    • 1
  • Tongxin Liu
    • 1
  • Yaqiong Zhang
    • 1
  • Xiaofang Liu
    • 1
  1. 1.State Key Laboratory for Agrobiotechnology, College of Biological SciencesChina Agricultural UniversityBeijingPeople’s Republic of China

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