Department of Biochemistry and Molecular Biophysics, Columbia University Medical Center, 701 West 168th Street, HHSC 1310, New York, NY, 10032, USA.
HHMI, Department of Biochemistry and Molecular Biophysics, and Department of Microbiology and Immunology, Columbia University Medical Center, New York, NY, 10032, USA.
Virol J. 2020 Feb 10;17(1):22. doi: 10.1186/s12985-020-1297-3.
Transfection, the process of introducing purified nucleic acids into cells, and viral transduction, viral-mediated nucleic acid transfer, are two commonly utilized techniques for gene delivery in the research setting. Transfection allows purified nucleic acid to be introduced into target cells through chemical-based techniques, nonchemical methods or particle-based methods, while viral transduction employs genomes or vectors based on adenoviruses, retroviruses (e.g. lentiviruses), adeno-associated viruses, or hybrid viruses. Transfected DNAs are often tested for potential effects on subsequent transduction, but it is not clear whether transfection itself rather than the particular nucleic acid being introduced might impact subsequent viral transfection. We observed a significant association between successfully transfected mobilized peripheral blood CD34+ human stem and progenitor cells (HSPCs) and permissiveness to subsequent lentiviral transduction, which was not evident in other cells such as 293 T cells and Jurkat cells. This association, apparently specific to CD34+ human stem and progenitor cells (HSPCs), is critical to both research and clinical applications as these cells are a frequent target of transfection and viral transduction owing to the durable nature of these cells in living systems. This finding may also present a significant opportunity to enhance the success of viral transduction for clinical applications.
转染是将纯化的核酸导入细胞的过程,而病毒转导是病毒介导的核酸转移,这两种技术是研究中常用的基因传递方法。转染允许通过基于化学的技术、非化学方法或基于粒子的方法将纯化的核酸导入靶细胞,而病毒转导则采用基于腺病毒、逆转录病毒(例如慢病毒)、腺相关病毒或杂交病毒的基因组或载体。转染的 DNA 通常会测试其对后续转导的潜在影响,但尚不清楚是转染本身而不是引入的特定核酸可能会影响随后的病毒转染。我们观察到成功转染的动员外周血 CD34+人类造血干/祖细胞 (HSPC) 与随后慢病毒转导的易感性之间存在显著关联,而在其他细胞(如 293T 细胞和 Jurkat 细胞)中则没有这种关联。这种关联显然是 CD34+人类造血干/祖细胞 (HSPC) 所特有的,这对于研究和临床应用都至关重要,因为由于这些细胞在活系统中的持久性,这些细胞是转染和病毒转导的常见靶标。这一发现也可能为提高病毒转导在临床应用中的成功率提供重要机会。