Pseudo typed baculovirus is an effective gene expression tool for studying
Pseudo typed baculovirus is an effective gene expression tool for studying
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DOI：
10.1016/j.ydbio.2017.10.008
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发表时间：
2018-01-15
影响因子：
2.7
通讯作者：
Tanaka, Elly M.
中科院分区：
文献类型：
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作者：
Oliveira, Catarina R.;Lemaitre, Regis;Tanaka, Elly M.
蝾螈可以通过成熟组织内细胞的募集和重塑来再生复杂的结构。以分子分辨率了解潜在机制对于了解损伤如何触发再生及其进行过程至关重要。然而，成人组织中的基因转化可能具有挑战性。在这里，我们描述了使用假型杆状病毒（BV）作为成熟肢体组织内和胚基内细胞的有效基因转移方法。这些细胞在转导后仍然能够参与再生。我们通过在芽基中过度表达Shh，进一步表征了BV对于基因过度表达研究的有效性，这产生了经典多指表型的高外显率。总体而言，我们的工作将 BV 确立为获取蝾螈肢体再生基因功能的强大工具。
Axolotls can regenerate complex structures through recruitment and remodeling of cells within mature tissues. Accessing the underlying mechanisms at a molecular resolution is crucial to understand how injury triggers regeneration and how it proceeds. However, gene transformation in adult tissues can be challenging. Here we characterize the use of pseudotyped baculovirus (BV) as an effective gene transfer method both for cells within mature limb tissue and within the blastema. These cells remain competent to participate in regeneration after transduction. We further characterize the effectiveness of BV for gene overexpression studies by overexpressing Shh in the blastema, which yields a high penetrance of classic polydactyly phenotypes. Overall, our work establishes BV as a powerful tool to access gene function in axolotl limb regeneration.