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Tion in any medium, provided the original work is properly cited. The Creative Commons Public Domain Dedication waiver (http:// creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.
Khattak et al. BMC Developmental Biology 2013, 13:17 http://www.biomedcentral.com/1471-213X/13/METHODOLOGY ARTICLEOpen AccessFoamy virus for efficient gene transfer in regeneration studiesShahryar Khattak1,2, Tatiana Sandoval-Guzm 1,2, Nicole Stanke2,3, Stephanie Protze1,2, Elly M Tanaka1,2* and Dirk Lindemann2,3*AbstractBackground: Molecular studies of appendage regeneration have been hindered by the lack of a stable and efficient means of transferring exogenous genes. We therefore sought an efficient integrating virus system that could be used to study limb and tail regeneration in salamanders. Results: We show that replication-deficient foamy virus (FV) vectors efficiently transduce cells in two different regeneration models in cell culture and in vivo. Injection of EGFP-expressing FV but not lentivirus vector particles into regenerating limbs and tail resulted in widespread expression that persisted throughout regeneration and reamputation pointing to the utility of FV for analyzing adult phenotypes in non-mammalian models. Furthermore, tissue specific transgene expression is achieved using FV vectors during limb regeneration. Conclusions: FV vectors are efficient mean of transferring genes into axolotl limb/tail and infection persists throughout regeneration and reamputation. This is a nontoxic method of delivering genes into axolotls in vivo/ in vitro and can potentially be applied to other salamander species. Keywords: Foamy virus, Regeneration, Salamander, in vivo gene transferBackground Limb regeneration of salamanders has fascinated scientists for several decades. Salamanders, Newts and Axolotls, are used as model animals in limb regeneration studies. Regeneration of missing structures is achieved by blastema, a pool of restricted progenitors that is formed after amputation [1]. Electroporation of DNA is the fastest and efficient method to introduce exogenous transgenes into salamander limb and spinal cord in vivo but this expression is lost during regeneration as the electroporated DNA is episomal [2]. Infection of cells both in cell culture and in vivo by modified LCZ696 chemical information viruses has been a powerful means of expressing exogenous genes in various experimental systems. For example, retroviral infection has been crucial for the molecular analysis of chicken limb development [3]. A* Correspondence: [email protected]; [email protected] de 1 Max Planck Institute of Molecular Cell Biology and Genetics, Pfotenhauerstr 108, 01307 Dresden, Germany 2 DFG-Center for RegenerativeTherapies Dresden, Technische Universit Dresden, Fetscherstr. 105, 01307 Dresden, Germany Full list of author information is available at the end of the articlecorresponding molecular analysis of limb regeneration in salamanders has been limited due to a paucity in gene transduction methods. Vaccinia and adenovirus have been used in limb regeneration studies, but their toxicity and non-integration phenotype limits their effectiveness [4,5]. PubMed ID:https://www.ncbi.nlm.nih.gov/pubmed/25957400 Similarly, pseudotyped retroviruses have been used in cultured cells [5-7] and a recent report showing their use in PubMed ID:https://www.ncbi.nlm.nih.gov/pubmed/26795252 vivo but the issue of viral silencing after second round of regeneration was not investigated [8]. We therefore sought a virus system that efficiently and st.

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