Tsubota Takuya, Sezutsu Hideki
Transgenic Silkworm Research Unit, Institute of Agrobiological Sciences, National Agriculture and Food Research Organization, 1-2 Owashi, Tsukuba, Ibaraki, 305-8634, Japan.
Methods Mol Biol. 2017;1630:205-218. doi: 10.1007/978-1-4939-7128-2_17.
Silkworm is a lepidopteran insect that has been used as a model for a wide variety of biological studies. The microinjection technique is available and it is possible to cause transgenesis as well as target gene disruption via the genome editing technique. TALEN-mediated knock-out is especially effective in this species. We also succeeded in the precise and efficient integration of a donor vector using the Precise Integration into Target Chromosome (PITCh) method. Here, we describe protocols for ZFN, TALEN, and CRISPR/Cas9-mediated genome editing as well as the PITCh technique in the silkworm. We consider that all of these techniques can contribute to the further promotion of various biological studies in the silkworm and other insect species.
家蚕是一种鳞翅目昆虫,已被用作多种生物学研究的模型。微注射技术可用,并且通过基因组编辑技术可以实现转基因以及靶基因破坏。TALEN介导的敲除在该物种中特别有效。我们还使用精确整合到靶染色体(PITCh)方法成功实现了供体载体的精确高效整合。在这里,我们描述了家蚕中ZFN、TALEN和CRISPR/Cas9介导的基因组编辑以及PITCh技术的方案。我们认为所有这些技术都有助于进一步推动家蚕和其他昆虫物种的各种生物学研究。