Receptor targeting of vector particles is a key technology to enable cell type–specific in vivo gene delivery. For example, T cells in humanized mouse models can be modified by lentiviral vectors (LVs) targeted to human T-cell markers to enable them to express chimeric antigen receptors (CARs). Here, we provide detailed protocols for the generation of CD4- and CD8-targeted LVs (which takes ~9 d in total). We also describe how to humanize immunodeficient mice with hematopoietic stem cells (which takes 12–16 weeks) and precondition (over 5 d) and administer the vector stocks. Conversion of the targeted cell type is monitored by PCR and flow cytometry of blood samples. A few weeks after administration, ~10% of the targeted T-cell subtype can be expected to have converted to CAR T cells. By closely following the protocol, sufficient vector stock for the genetic manipulation of 10–15 humanized mice is obtained. We also discuss how the protocol can be easily adapted to use LVs targeted to other types of receptors and/or for delivery of other genes of interest.

载体颗粒的受体靶向是实现细胞类型特异性体内基因递送的关键技术。例如，人源化小鼠模型中的 T 细胞可以通过靶向人类 T 细胞标志物的慢病毒载体 (LV) 进行修饰，使其能够表达嵌合抗原受体 (CAR)。在这里，我们提供了用于生成 CD4 和 CD8 目标 LV 的详细协议（总共需要约 9 天）。我们还描述了如何用造血干细胞（需要 12-16 周）和预处理（超过 5 天）对免疫缺陷小鼠进行人源化，并管理载体储备。通过血液样本的 PCR 和流式细胞术监测目标细胞类型的转化。给药几周后，预计约 10% 的靶向 T 细胞亚型已转化为 CAR T 细胞。通过严格遵守协议，获得了足够的载体库，用于对 10-15 只人源化小鼠进行基因操作。我们还讨论了如何轻松调整该协议以使用针对其他类型受体和/或传递其他感兴趣基因的 LV。