Wellcome Trust Sanger Institute, Genome Campus, Hinxton-Cambridge CB10 1SA, UK.
Science. 2010 Nov 19;330(6007):1104-7. doi: 10.1126/science.1193004. Epub 2010 Oct 14.
Transposons are mobile DNA segments that can disrupt gene function by inserting in or near genes. Here, we show that insertional mutagenesis by the PiggyBac transposon can be used for cancer gene discovery in mice. PiggyBac transposition in genetically engineered transposon-transposase mice induced cancers whose type (hematopoietic versus solid) and latency were dependent on the regulatory elements introduced into transposons. Analysis of 63 hematopoietic tumors revealed that PiggyBac is capable of genome-wide mutagenesis. The PiggyBac screen uncovered many cancer genes not identified in previous retroviral or Sleeping Beauty transposon screens, including Spic, which encodes a PU.1-related transcription factor, and Hdac7, a histone deacetylase gene. PiggyBac and Sleeping Beauty have different integration preferences. To maximize the utility of the tool, we engineered 21 mouse lines to be compatible with both transposon systems in constitutive, tissue- or temporal-specific mutagenesis. Mice with different transposon types, copy numbers, and chromosomal locations support wide applicability.
转座子是可移动的 DNA 片段，可通过插入基因或基因附近而破坏基因功能。在这里，我们表明，PiggyBac 转座子的插入诱变可用于在小鼠中发现癌症基因。在基因工程转座子-转座酶小鼠中，PiggyBac 转座诱导的癌症类型（造血性与实体性）和潜伏期取决于引入转座子的调节元件。对 63 种造血肿瘤的分析表明，PiggyBac 能够进行全基因组诱变。PiggyBac 筛选揭示了许多以前在逆转录病毒或 Sleeping Beauty 转座子筛选中未鉴定出的癌症基因，包括编码与 PU.1 相关的转录因子 Spic 和组蛋白去乙酰化酶基因 Hdac7。PiggyBac 和 Sleeping Beauty 具有不同的整合偏好。为了最大化工具的效用，我们设计了 21 条小鼠品系，以在组成型、组织特异性或时间特异性诱变中与两种转座子系统兼容。具有不同转座子类型、拷贝数和染色体位置的小鼠支持广泛的适用性。
