Kuo Tzu-Hsing, Whited Jessica L
Brigham Regenerative Medicine Center, Department of Orthopedic Surgery, Brigham & Women's Hospital, Cambridge, MA, USA.
Methods Mol Biol. 2015;1290:127-40. doi: 10.1007/978-1-4939-2495-0_10.
The ability to introduce DNA elements into host cells and analyze the effects has revolutionized modern biology. Here we describe a protocol to generate Moloney murine leukemia virus (MMLV)-based, replication-incompetent pseudotyped retrovirus capable of infecting axolotls and incorporating genetic information into their genome. When pseudotyped with vesicular stomatitis virus (VSV)-G glycoprotein, the retroviruses can infect a broad range of proliferative axolotl cell types. However, if the retrovirus is pseudotyped with an avian sarcoma leukosis virus (ASLV)-A envelope protein, only axolotl cells experimentally manipulated to express the cognate tumor virus A (TVA) receptor can be targeted by infections. These strategies enable robust transgene expression over many cell divisions, cell lineage tracing, and cell subtype targeting for gene expression.
将DNA元件导入宿主细胞并分析其效果的能力彻底改变了现代生物学。在此，我们描述了一种方案，用于生成基于莫洛尼鼠白血病病毒（MMLV）的、无复制能力的假型逆转录病毒，该病毒能够感染蝾螈并将遗传信息整合到它们的基因组中。当用水泡性口炎病毒（VSV）-G糖蛋白进行假型化时，逆转录病毒可以感染多种增殖性蝾螈细胞类型。然而，如果逆转录病毒用禽肉瘤白血病病毒（ASLV）-A包膜蛋白进行假型化，那么只有经过实验操作以表达同源肿瘤病毒A（TVA）受体的蝾螈细胞才能被感染靶向。这些策略能够在许多细胞分裂过程中实现强大的转基因表达、细胞谱系追踪以及针对基因表达的细胞亚型靶向。
