Journal article
Robust and ubiquitous GFP expression in a single generation of chicken embryos using the avian retroviral vector, RCASBP
CA Smith, KN Roeszler, AH Sinclair
Differentiation | ELSEVIER SCI LTD | Published : 2009
Abstract
Functional genomics in avian models has lagged behind that of mammals, and the production of transgenic birds has proven to be challenging and time-consuming. All current methods rely upon breeding chimeric birds through at least one generation. Here, we report a rapid method for the ubiquitous expression of GFP in chicken embryos in a single generation (G-0), using the avian retroviral vector, Replication-Competent Avian sarcoma-leukosis virus, with a Splice acceptor, Bryan RSV Pol (RCASBP). High-titre RCASBP retrovirus carrying eGFP was injected into unincubated (stage X) blastoderms in ovo. This resulted in stable and widespread expression of eGFP throughout development in a very high pro..
View full abstractGrants
Funding Acknowledgements
We thank Dr. Helen Sang, Dr. Simon Lilico, Dr. Mike Clinton and Dr. Mike McGrew at the Roslin Institute, the United Kingdom, for demonstrating viral injections and embryomanipulation. Professor Cliff Tabin (Harvard Medical School) provided the viral vectors and Dr. Richard Pearce (Harvard Medical School) provided helpful advice. This work was supported by an Australian Research Council grant to CAS and AHS.