Department of Biomedicine, Aarhus University, Aarhus, Denmark.
Mol Ther Nucleic Acids. 2013 Feb 26;2(2):e74. doi: 10.1038/mtna.2013.1.
DNA transposon-based vectors have emerged as new potential delivery tools in therapeutic gene transfer. Such vectors are now showing promise in hematopoietic stem cells and primary human T cells, and clinical trials with transposon-engineered cells are on the way. However, the use of plasmid DNA as a carrier of the vector raises safety concerns due to the undesirable administration of bacterial sequences. To optimize vectors based on the Sleeping Beauty (SB) DNA transposon for clinical use, we examine here SB transposition from DNA minicircles (MCs) devoid of the bacterial plasmid backbone. Potent DNA transposition, directed by the hyperactive SB100X transposase, is demonstrated from MC donors, and the stable transfection rate is significantly enhanced by expressing the SB100X transposase from MCs. The stable transfection rate is inversely related to the size of circular donor, suggesting that a MC-based SB transposition system benefits primarily from an increased cellular uptake and/or enhanced expression which can be observed with DNA MCs. DNA transposon and transposase MCs are easily produced, are favorable in size, do not carry irrelevant DNA, and are robust substrates for DNA transposition. In accordance, DNA MCs should become a standard source of DNA transposons not only in therapeutic settings but also in the daily use of the SB system.Molecular Therapy - Nucleic Acids (2013) 2, e74; doi:10.1038/mtna.2013.1; published online 26 February 2013.
基于 DNA 转座子的载体已成为治疗性基因转移的新的潜在递药工具。此类载体目前在造血干细胞和原代人 T 细胞中显示出前景，采用转座子工程化细胞的临床试验正在进行中。然而，由于细菌序列的不期望的给药，将质粒 DNA 用作载体可能会引发安全性问题。为了优化基于 Sleeping Beauty（SB）DNA 转座子的载体用于临床应用，我们在此检查了不含细菌质粒骨架的 DNA 微环（MC）中的 SB 转座。从 MC 供体中证明了由超活性 SB100X 转座酶指导的有效的 DNA 转座，并且通过从 MC 表达 SB100X 转座酶来显著增强稳定转染率。稳定转染率与环状供体的大小呈反比，这表明基于 MC 的 SB 转座系统主要受益于增加的细胞摄取和/或可观察到的 DNA MC 中增强的表达。DNA 转座子和转座酶 MC 易于制备，大小有利，不携带无关 DNA，并且是 DNA 转座的稳健底物。因此，DNA MC 不仅应成为治疗性设置中，而且还应成为 SB 系统日常使用中的 DNA 转座子的标准来源。
Molecular Therapy - Nucleic Acids (2013) 2, e74; doi:10.1038/mtna.2013.1; published online 26 February 2013.
