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首页> 外文期刊>Journal of insect biotechnology and sericology >Enlargement of egg size by CRISPR/Cas9-mediated knockout of a sex-linked gene in the silkworm, Bombyx mori
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Enlargement of egg size by CRISPR/Cas9-mediated knockout of a sex-linked gene in the silkworm, Bombyx mori

机译：通过CRISPR / CAS9介导的鸡蛋大小的敲击鸡蛋蚕的爆击鸡肉中的鸡蛋，Bombyx Mori

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摘要

Ge (giant egg) is a sex-linked mutation in Bombyx mori which leads to the production of large-sized eggs caused by an increase in the number of follicular epithelial cells, each with decreased cell size. We previously determined that a gene orthologous to human Phytanoyl-CoA dioxygenase domain-containing 1 (PHYHD1) is specifically disrupted in Ge and Ge2 mutants through positional cloning of the Ge locus. However, little is known about the function of the BmPHYHDI gene in determination of the egg size. In this study, we knocked out BmPHYHDI using CRISPR/Cas9. We employed a novel knockout protocol developed in other lepidopteran species in which a commercialized Cas9 mixed with two sgRNAs designed for a target gene is injected into newly-laid eggs. First, we confirmed that the protocol was applicable to B. mori with high efficiency. Second, we targeted the BmPHYHDI gene and obtained G0 moths laid large size eggs. CRISPR/Cas9-mediated mutations were confirmed in G, moths that laid large-sized eggs, demonstrating that the BmPHYHDI gene controls egg size in 6. morì. Finally, the dominant/recessive relationship between Ge and +Ge is discussed and we propose the use of gi (giant egg) and gi2 (giant egg 2) to represent giant egg mutations, Ge and Ge2, respectively. We believe that Ge mutants are an appropriate model for the study of genetic mechanisms mediating the size and number of cells in animals.
机译：GE（巨蛋）是Bombyx Mori中的性关系突变，导致大型卵产生由滤泡上皮细胞数量的增加，每个卵泡细胞的数量降低。之前，我们以通过鉴于GE基因座的定位克隆，在GE和GE2突变体中明确地中断了对含人植物植物植物植物 - 辅酶二氧基酶的1（PHYHD1）的基因。然而，关于Bmphyhdi基因在测定蛋尺寸时几乎熟知。在这项研究中，我们使用CRISPR / CAS9淘汰了BMPHYHDI。我们雇用了一种在其他鳞翅目种类中开发的新型敲除协议，其中将与为靶基因设计的两个SGRNA混合的商业化CAS9注入新占鸡蛋中。首先，我们确认该协议适用于B. Mori，效率高。其次，我们瞄准了BMPhyhdi基因，并获得了G0蛾奠定了大型鸡蛋。 CRISPR / CAS9介导的突变在G，蛾的蛾中确认，铺设了大型卵子，表明BMPhyhdi基因在6.Morì中对照卵子尺寸。最后，讨论了GE和+ GE之间的主导/隐性关系，并提出了使用GI（巨蛋）和GI2（巨蛋2）分别代表巨蛋突变，GE和GE2。我们认为GE突变体是研究介导动物中细胞的大小和数量的遗传机制的适当模型。

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    Tsuguru Fujii; Yutaka Banno;

  • 作者单位

    Silkworm Resource Division Institute of Genetic Resources Kyushu University Graduate School of Bioresources and Bioenvironmental Science 744 Motooka Nishi-ku Fukuoka 819-0395 Japan;

    Silkworm Resource Division Institute of Genetic Resources Kyushu University Graduate School of Bioresources and Bioenvironmental Science 744 Motooka Nishi-ku Fukuoka 819-0395 Japan;

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  • 原文格式 PDF
  • 正文语种 eng
  • 中图分类 蚕桑;
  • 关键词

    oogenesis; choriogenesis; uric acid; allometry; genome editing;

    机译：ofocoisesis;核发生;尿酸;almometry;基因组编辑;

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