title: pTransgenesis: a cross-species, modular transgenesis resource.
abstract: As studies aim increasingly to understand key, evolutionarily conserved properties of biological systems, the ability to move transgenesis experiments efficiently between organisms becomes essential. DNA constructions used in transgenesis usually contain four elements, including sequences that facilitate transgene genome integration, a selectable marker and promoter elements driving a coding gene. Linking these four elements in a DNA construction, however, can be a rate-limiting step in the design and creation of transgenic organisms. In order to expedite the construction process and to facilitate cross-species collaborations, we have incorporated the four common elements of transgenesis into a modular, recombination-based cloning system called pTransgenesis. Within this framework, we created a library of useful coding sequences, such as various fluorescent protein, Gal4, Cre-recombinase and dominant-negative receptor constructs, which are designed to be coupled to modular, species-compatible selectable markers, promoters and transgenesis facilitation sequences. Using pTransgenesis in Xenopus, we demonstrate Gal4-UAS binary expression, Cre-loxP-mediated fate-mapping and the establishment of novel, tissue-specific transgenic lines. Importantly, we show that the pTransgenesis resource is also compatible with transgenesis in Drosophila, zebrafish and mammalian cell models. Thus, the pTransgenesis resource fosters a cross-model standardization of commonly used transgenesis elements, streamlines DNA construct creation and facilitates collaboration between researchers working on different model organisms.
authors: Faculty of Life Sciences, University of Manchester, Oxford Road, Manchester, UK.
authors: Nick R
authors: NR
authors: Love
authors: Raphael
authors: R
authors: Thuret
authors: Yaoyao
authors: Y
authors: Chen
authors: Shoko
authors: S
authors: Ishibashi
authors: Nitin
authors: N
authors: Sabherwal
authors: Roberto
authors: R
authors: Paredes
authors: Juliana
authors: J
authors: Alves-Silva
authors: Karel
authors: K
authors: Dorey
authors: Anna M
authors: AM
authors: Noble
authors: Matthew J
authors: MJ
authors: Guille
authors: Yoshiki
authors: Y
authors: Sasai
authors: Nancy
authors: N
authors: Papalopulu
authors: Enrique
authors: E
authors: Amaya
mesh_terms: D000818:Animals; D030801:Animals, Genetically Modified / Q000235:genetics*; D004330:Drosophila / Q000235:genetics; D015723:Gene Library; D018014:Gene Transfer Techniques; D019426:Integrases / Q000378:metabolism; D014157:Transcription Factors / Q000235:genetics; D019076:Transgenes; D014981:Xenopus / Q000235:genetics; D015027:Zebrafish / Q000235:genetics
chemical_list: D014157:Transcription Factors; C045073:Cre recombinase; D019426:Integrases