Wellcome Trust Sanger Institute, Wellcome Trust Genome Campus, Hinxton, Cambridge, UK, CB10 1SA.
Nucleic Acids Res. 2011 Dec;39(22):e148. doi: 10.1093/nar/gkr764. Epub 2011 Sep 24.
The development of technologies that allow the stable delivery of large genomic DNA fragments in mammalian systems is important for genetic studies as well as for applications in gene therapy. DNA transposons have emerged as flexible and efficient molecular vehicles to mediate stable cargo transfer. However, the ability to carry DNA fragments >10 kb is limited in most DNA transposons. Here, we show that the DNA transposon piggyBac can mobilize 100-kb DNA fragments in mouse embryonic stem (ES) cells, making it the only known transposon with such a large cargo capacity. The integrity of the cargo is maintained during transposition, the copy number can be controlled and the inserted giant transposons express the genomic cargo. Furthermore, these 100-kb transposons can also be excised from the genome without leaving a footprint. The development of piggyBac as a large cargo vector will facilitate a wider range of genetic and genomic applications.
在哺乳动物系统中稳定递增大基因组 DNA 片段的技术的发展对于遗传研究以及基因治疗中的应用都很重要。DNA 转座子已成为介导稳定载体转移的灵活有效的分子载体。然而，大多数 DNA 转座子携带 DNA 片段的能力>10kb 是有限的。在这里，我们表明 DNA 转座子 piggyBac 可以在小鼠胚胎干细胞（ES）细胞中移动 100kb 的 DNA 片段，使其成为唯一已知具有如此大容量载物的转座子。在转座过程中保持货物的完整性，可以控制拷贝数，插入的巨型转座子表达基因组货物。此外，这些 100kb 的转座子也可以从基因组中切除而不会留下痕迹。piggyBac 作为一个大的货物载体的发展将促进更广泛的遗传和基因组应用。
