Li Ming-Jie, Rossi John J
CSH Protoc. 2007 May 1;2007:pdb.prot4755. doi: 10.1101/pdb.prot4755.
INTRODUCTIONEfficient transfer and sustained expression of transgenes are among the most important issues in gene delivery. The majority of hematopoietic cells are nondividing or slowly self-renewing. Thus, they are refractory to most nonviral or retroviral delivery methods. Lentiviral vectors are capable of transducing nondividing cells and maintaining long-term and sustained expression of the transgenes. They are becoming useful for many delivery protocols, such as long-term expression of short hairpin RNA (shRNA) and functional genetics. They may also have great potential in gene therapy. This protocol describes lentivirus-vector-based delivery of foreign genes to hematopoietic cells. The method is applicable to various cell types in experiments that require long-term transgene expression.
引言
转基因的高效转移和持续表达是基因递送中最重要的问题之一。大多数造血细胞不进行分裂或自我更新缓慢。因此,它们对大多数非病毒或逆转录病毒递送方法具有抗性。慢病毒载体能够转导非分裂细胞并维持转基因的长期持续表达。它们在许多递送方案中变得很有用,例如短发夹RNA(shRNA)的长期表达和功能遗传学。它们在基因治疗中也可能具有巨大潜力。本方案描述了基于慢病毒载体将外源基因递送至造血细胞的方法。该方法适用于需要长期转基因表达的实验中的各种细胞类型。