Khattak Shahryar, Sandoval-Guzmán Tatiana, Stanke Nicole, Protze Stephanie, Tanaka Elly M, Lindemann Dirk
Max Planck Institute of Molecular Cell Biology and Genetics, Pfotenhauerstr 108, 01307 Dresden, Germany.
BMC Dev Biol. 2013 May 3;13:17. doi: 10.1186/1471-213X-13-17.
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.
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.
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.
由于缺乏稳定、高效的外源基因转移方法，附肢再生的分子研究受到了阻碍。因此，我们寻求一种高效的整合病毒系统，用于研究蝾螈的肢体和尾巴再生。
我们发现，复制缺陷型泡沫病毒（FV）载体能在细胞培养和体内的两种不同再生模型中高效转导细胞。将表达绿色荧光蛋白（EGFP）的FV载体颗粒而非慢病毒载体颗粒注射到再生的肢体和尾巴中，会导致广泛表达，这种表达在整个再生和再次截肢过程中持续存在，这表明FV在分析非哺乳动物模型中的成体表型方面具有实用性。此外，在肢体再生过程中使用FV载体可实现组织特异性转基因表达。
FV载体是将基因导入蝾螈肢体/尾巴的有效手段，并且感染在整个再生和再次截肢过程中持续存在。这是一种在体内/体外将基因导入蝾螈的无毒方法，并且有可能应用于其他蝾螈物种。
