RNA Interference in Chicken Embryos

  • Nick J. Van Hateren
  • Rachel S. Jones
  • Stuart A. Wilson

The chicken has played an important role in biological discoveries since the 17th century (Stern, 2005). Many investigations into vertebrate development have utilized the chicken due to the accessibility of the chick embryo and its ease of manipulation (Brown et al., 2003). However, the lack of genetic resources has often handicapped these studies and so the chick is frequently overlooked as a model organism for the analysis of vertebrate gene function in favor of mice or zebrafish. In the past six years this situation has altered dramatically with the generation of over half a million expressed sequence tags and >20,000 fully sequenced chicken cDNAs (Boardman et al. 2002; Caldwell et al., 2005; Hubbard et al., 2005) together with a 6X coverage genome sequence (Hillier et al., 2004). These resources have created a comprehensive catalogue of chicken genes with readily accessible cDNA and EST resources available via ARK-GENOMICS ( for the functional analysis of vertebrate gene function.

The chicken embryo is conveniently packaged in an egg shell and it is a relatively straightforward process to create a window in this shell. This allows access to the embryo at any stage of development and facilitates manipulation of the embryo. After this procedure, the window can be resealed and the egg incubated for a suitable time period prior to analyzing the results of the manipulation. Such manipulations traditionally involved ”cut and paste“ experiments in which tissue is excised and transplanted to ectopic locations in the embryo or from quail embryos, which are readily distinguished histologically, to chick embryos to generate chimeras. Whilst these studies have led to many important discoveries (Brown et al., 2003), there has recently been an increase in direct genetic manipulation approaches that can be applied to the chick embryo.


Neural Tube Chick Embryo Chicken Embryo RNAi Vector Prehybridization Solution 
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  1. Albazerchi A, Cinquin O, Stern CD (2007) A new method to transfect the hypoblast of the chick embryo reveals conservation of the regulation of an Otx2 enhancer between mouse and chick extra embryonic endoderm. BMC Dev Biol 7: 25.CrossRefGoogle Scholar
  2. Boardman PE, Sanz-Ezquerro J, Overton IM, Burt D, Bosch E, Fong W, Tickle C, Brown WRA, Wilson SA, Hubbard SJ (2002) A comprehensive collection of chicken cDNAs. Curr Biol 12: 1965–1969.CrossRefGoogle Scholar
  3. Bron R, Eickholt BJ, Vermeren M, et al. (2004) Functional knockdown of neuropilin-1 in the developing chick nervous system by siRNA hairpins phenocopies genetic ablation in the mouse. Dev Dyn 230: 299–308.CrossRefGoogle Scholar
  4. Brown WR, Hubbard SJ, Tickle C, Wilson, SA (2003) The chicken as a model for large-scale analysis of vertebrate gene function. Nat Rev Genet 4: 87–98.CrossRefGoogle Scholar
  5. Caldwell RB, Kierzek AM, Arakawa H, et al. (2005) Full-length cDNAs from chicken bursal lymphocytes to facilitate gene function analysis. Genome Biol 6: R6.CrossRefGoogle Scholar
  6. Chesnutt C, Niswander L (2004) Plasmid-based short-hairpin RNA interference in the chicken embryo. Genesis 39: 73–78.CrossRefGoogle Scholar
  7. Dai F, Yusuf F, Farjah GH, et al. (2005) RNAi-induced targeted silencing of developmental control genes during chicken embryogenesis. Dev Biol 285: 80–90.CrossRefGoogle Scholar
  8. Das RM, Van Hateren NJ, Howell GR, et al. (2006) A robust system for RNA interference in the chicken using a modified microRNA operon. Dev Biol 294: 554–563.CrossRefGoogle Scholar
  9. Elbashir SM, Harborth J, Lendeckel W, et al. (2001) Duplexes of 21-nucleotide RNAs mediate RNA interference in cultured mammalian cells. Nature 411: 494–498.CrossRefGoogle Scholar
  10. Hamburger V, Hamilton HL (1951) A series of normal stages in the development of the chick embryo. J. Morphol 88: 49–92.CrossRefGoogle Scholar
  11. Harpavat S, Cepko CL (2006) RCAS-RNAi: a loss-of-function method for the developing chick retina. BMC Dev Biol 6: 2.CrossRefGoogle Scholar
  12. Hillier L, Miller W, Birney E, et al. (2004) Sequence and comparative analysis of the chicken genome provide unique perspectives on vertebrate evolution. Nature 432: 695–716.CrossRefGoogle Scholar
  13. Hu WY, Myers CP, Kilzer JM, et al. (2002) Inhibition of retroviral pathogenesis by RNA interference. Curr Biol 12: 1301–1311.CrossRefGoogle Scholar
  14. Hubbard SJ, Grafham DV, Beattie KJ, et al. (2005) Transcriptome analysis for the chicken based on 19,626 finished cDNA sequences and 485,337 expressed sequence tags. Genome Res 15: 174–183.CrossRefGoogle Scholar
  15. Isaacs A, Lindenmann J (1957) Virus interference. I. The interferon. Proc R Soc Lond B Biol Sci 147: 258–267.CrossRefGoogle Scholar
  16. Itasaki N, Bel-Vialar S, Krumlauf R (1999) ‘Shocking’ developments in chick embryology: elec-troporation and in ovo gene expression. Nat Cell Biol 1: E203–207.CrossRefGoogle Scholar
  17. Katahira T, Nakamura H (2003) Gene silencing in chick embryos with a vector-based small interfering RNA system. Dev Growth Differ 45: 361–367.CrossRefGoogle Scholar
  18. Kudo T, Sutou S (2005) Usage of putative chicken U6 promoters for vector-based RNA interference. J Reprod Dev. 51: 411–417.CrossRefGoogle Scholar
  19. Ohta S, Suzuki K, Tachibana K, et al. (2003) Microbubble-enhanced sonoporation: efficient gene transduction technique for chick embryos. Genesis 37: 91–101.CrossRefGoogle Scholar
  20. Pekarik V, Bourikas D, Miglino N, et al. (2003) Screening for gene function in chicken embryo using RNAi and electroporation. Nat Biotechnol 21: 93–96.CrossRefGoogle Scholar
  21. Shiau CE, Lwigale P Y, Das RM, Wilson SA, Bronner-Fraser M (2008) Robo2-Slit1 dependent cell–cell interactions mediate assembly of the trigeminal ganglion. Nat Neurosci 11: 269–76.CrossRefGoogle Scholar
  22. Stern CD (2005) The chick; a great model system becomes even greater. Dev Cell 8: 9–17.Google Scholar
  23. Wong GK, Liu B, Wang J, et al. (2004) A genetic variation map for chicken with 2.8 million single-nucleotide polymorphisms. Nature 432: 717–722.CrossRefGoogle Scholar
  24. Zeng Y, Cullen BR (2005) Efficient processing of primary microRNA hairpins by Drosha requires flanking nonstructured RNA sequences. J Biol Chem 280: 27595–27603.CrossRefGoogle Scholar

Copyright information

© Springer 2009

Authors and Affiliations

  • Nick J. Van Hateren
    • 1
  • Rachel S. Jones
    • 1
  • Stuart A. Wilson
    • 1
  1. 1.Department of Molecular Biology and BiotechnologyThe University of SheffieldSheffieldUK

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