Institute for Molecular Medicine, University Medical Center of the Johannes Gutenberg-University Mainz, 55131 Mainz, Germany.
Eur J Immunol. 2015 Jun;45(6):1596-9. doi: 10.1002/eji.201545685. Epub 2015 May 12.
Ectopic gene expression studies in primary immune cells have been notoriously difficult to perform due to the limitations in conventional transfection and viral transduction methods. Although replication-defective adenoviruses provide an attractive alternative for gene delivery, their use has been hampered by the limited susceptibility of murine leukocytes to adenoviral infection, due to insufficient expression of the human coxsackie/adenovirus receptor (CAR). In this issue of the European Journal of Immunology, Heger et al. [Eur. J. Immunol. 2015. 45: XXXX-XXXX] report the generation of transgenic mice that enable conditional Cre/loxP-mediated expression of human CAR. The authors demonstrate that this R26/CAG-CAR∆1(StopF) mouse strain facilitates the faithful monitoring of Cre activity in situ as well as the specific and efficient adenoviral transduction of primary immune cell populations in vitro. Further tweaking of the system towards more efficient gene transfer in vivo remains a future challenge.
由于传统的转染和病毒转导方法存在局限性,原代免疫细胞中的异位基因表达研究一直极具挑战性。虽然复制缺陷型腺病毒为基因传递提供了一种有吸引力的替代方法,但由于人类柯萨奇/腺病毒受体 (CAR) 的表达不足,鼠类白细胞对腺病毒感染的敏感性有限,其应用受到了阻碍。在本期的《欧洲免疫学杂志》中,Heger 等人[Eur. J. Immunol. 2015. 45: XXXX-XXXX]报道了生成转基因小鼠的方法,使人类 CAR 能够进行条件性 Cre/loxP 介导的表达。作者证明,这种 R26/CAG-CAR∆1(StopF) 小鼠品系可方便地原位监测 Cre 活性,以及体外原代免疫细胞群体的特异性和高效腺病毒转导。进一步调整该系统以实现更有效的体内基因转移仍然是未来的挑战。