Martinet Wim, Schrijvers Dorien M, Kockx Mark M
Division of Pharmacology, University of Antwerp, Wilrijk, Belgium.
Biotechnol Lett. 2003 Jul;25(13):1025-9. doi: 10.1023/a:1024157508492.
Despite some progress in the field of gene transfer into hard-to-transfect cells, so far an efficient nonviral method for monocytes has not been available. A comparison of plasmid DNA with capped and polyadenylated mRNA for enhanced green fluorescent protein gene delivery into the commonly used monocytic cell lines U937 and THP-1 suggested that limited DNA trafficking may be the underlying cause of poor transfection results. As Nucleofector technology delivers DNA (or mRNA) straight into the nucleus, we obtained nucleofection efficiencies of up to 80% without significant cell toxicity. Moreover, as the DNA quickly reaches the nucleus, nucleofected cells were ready for analysis after only 2-6 h. The technique is suitable not only for monocytes but also for other hard-to-transfect cells.
尽管在将基因导入难以转染的细胞领域取得了一些进展，但到目前为止，尚未有针对单核细胞的高效非病毒方法。将质粒DNA与加帽和聚腺苷酸化的mRNA用于增强型绿色荧光蛋白基因导入常用单核细胞系U937和THP-1的比较表明，有限的DNA转运可能是转染效果不佳的根本原因。由于核转染技术可将DNA（或mRNA）直接送入细胞核，我们获得了高达80%的核转染效率，且无明显细胞毒性。此外，由于DNA能迅速到达细胞核，经核转染的细胞仅需2 - 6小时即可进行分析。该技术不仅适用于单核细胞，也适用于其他难以转染的细胞。
