Status of transgenic chicken models for developmental biology

Mozdziak, Paul; Petitte, James N.

doi:10.1002/dvdy.10461

Cited by 75 publications

(63 citation statements)

References 42 publications

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“…The production of transgenic chickens has wide-ranging applications in academic research, biotechnology, and agriculture (51)(52)(53). Transgenic birds are model organisms for the study of developmental biology, as bioreactors for the production of therapeutic proteins, and as models of disease resistance to minimize losses in agriculture.…”

Section: Discussionmentioning

confidence: 99%

Efficient genetic modification and germ-line transmission of primordial germ cells using piggyBac and Tol2 transposons

Macdonald

Taylor

Sherman

et al. 2012

Proc. Natl. Acad. Sci. U.S.A.

155

127

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The derivation of germ-line competent avian primordial germ cells establishes a cell-based model system for the investigation of germ cell differentiation and the production of genetically modified animals. Current methods to modify primordial germ cells using DNA or retroviral vectors are inefficient and prone to epigenetic silencing. Here, we validate the use of transposable elements for the genetic manipulation of primordial germ cells. We demonstrate that chicken primordial germ cells can be modified in vitro using transposable elements. Both piggyBac and Tol2 transposons efficiently transpose primordial germ cells. Tol2 transposon integration sites were spread throughout both the macro-and microchromosomes of the chicken genome and were more prevalent in gene transcriptional units and intronic regions, consistent with transposon integrations observed in other species. We determined that the presence of insulator elements was not required for reporter gene expression from the integrated transposon. We further demonstrate that a gene-trap cassette carried in the Tol2 transposon can trap and mutate endogenous transcripts in primordial germ cells. Finally, we observed that modified primordial germ cells form functional gametes as demonstrated by the generation of transgenic offspring that correctly expressed a reporter gene carried in the transposon. Transposable elements are therefore efficient vectors for the genetic manipulation of primordial germ cells and the chicken genome.poultry | transgenesis | transposition P rimordial germ cells (PGCs) are specified in the early embryo and are the lineage-restricted stem cells for the germ cell population. In the chicken, segregation of PGCs from somatic cells occurs during the early stages of blastoderm formation (1-3). These cells accumulate in the germinal crescent region anterior to the forming head at day 1 of incubation [stage 4 Hamburger and Hamilton (HH)] and subsequently migrate via the circulatory system to the forming gonads on day 3 of incubation (stage 15 HH) (4). Chicken PGCs can be isolated from embryonic blood at this developmental stage and propagated indefinitely in culture (5). When returned to the circulatory system of an equivalent stage host embryo, the cultured PGCs migrated to and populated the forming embryonic gonad. Breeding from the resulting germline chimeras demonstrated that the cultured PGCs formed functional gametes by production of offspring derived from these cells. Germ-line transmission of cultured PGCs has now been demonstrated for three different chicken breeds (5-7).This in vitro system for the long-term propagation of chicken PGCs is the basis of a unique stem cell model for both the investigation of germ cell differentiation and the generation of genetically modified chickens. Thus far, chicken PGCs have proved highly resistant to genetic modification. Stable transfection of PGCs has been achieved by electroporation of plasmid DNA, at a frequency of ∼1 in 10 6 , but expression from integrated reporter constructs depended on t...

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Section: Discussionmentioning

confidence: 99%

Efficient genetic modification and germ-line transmission of primordial germ cells using piggyBac and Tol2 transposons

Macdonald

Taylor

Sherman

et al. 2012

Proc. Natl. Acad. Sci. U.S.A.

155

127

View full text Add to dashboard Cite

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“…Punctate staining was observed at variable locations on the skin of hatched geckos, but was never observed in the skin of negative control geckos. Mosaic gene expression and insertion is expected in the G0 founder animals (Mozdziak and Petitte, 2004) making the observation unsurprising that the first round of PCR amplification was not robust. In G0 animals, the lentiviral construct does not likely infect every cell in the embryo.…”

Section: Discussionmentioning

confidence: 99%

“…Furthermore, transgenic reptiles carrying reporter genes are useful tools for developmental biologists because the cells can be used to study the mechanisms of cell migration, differentiation, and cell fate in the same way that transgenic chickens can be employed to learn about avian development (Mozdziak and Petitte, 2004). The present results will unlock a new range of technology that can be employed to further understand the reptilian system.…”

Section: Discussionmentioning

confidence: 99%

Lentiviral transgenesis of the leopard gecko, Eublepharis macularius

Hull¹,

Hodgson²,

Clark³

et al. 2014

Afr. J. Microbiol. Res.

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Lentiviral vectors are an effective method of introducing transgenes into the genome of early stage embryos because they transduce both dividing and non-dividing cells. Lentiviral pseudoparticles containing the coding sequence for the fluorescent protein DsRed were injected into freshly laid leopard gecko eggs. Tissue samples were collected from hatchlings, and the samples were tested for the presence of the transgene. Of the injected gecko population, greater than 89% of efficiency of transgenesis was confirmed using polymerase chain reaction (PCR). Histological evaluations revealed the presence of DsRed 2 in injected gecko organs; with protein production concentrated in the muscle, kidney, and heart. Therefore, lentiviral vectors appear to be viable technology to create transgenic geckos.

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“…More interdisciplinary studies on model organisms representing animals in various branches of the phylogenetic tree will enhance our comparative study. (Berghammer et al, 1999;Chang et al, 2011;Dunham et al, 2002;Gonzalez-Estevez et al, 2003;Hong et al, 2009;Houdebine, 2009;Huss et al, 2008;Lu et al 1996;Macha et al, 1997;Mozdziak & Petitte 2004 ;Pavlopoulos and Averof, 2005;Renfer et al, 2010;Sasaki et al, 2009;Sasakura et al, 2007;Wheeler, 2003). The development of genome manipulation techniques for these emerging animal models will open the door to unlimited possibilities of in vivo investigations.…”

Section: Chilling Injurymentioning

confidence: 99%