Transformation of Gonipterus sp. n. 2 embryos with the CRISPR/Cas9 gene editing tool

Literature review discusses the insect sex determination pathway and CRISPR/Cas9 gene editing tool in application for genetic-based pest control methods. The de novo genome of Gonipterus sp. n. 2 was sequenced with long and short read sequencing, assembled and annotated using RNA sequencing data and Protein sequences from other Coleopteran species with published, assembled genomes. The expansion and contraction of gene families, with a focus on the cytochrome P450 detoxification gene, was assessed. The ground work was laid for a CRISPR/Cas9 transformation protocol for Gonipterus sp. n. 2 embryos with microinjection. The Tyrosine 3-monooxygenase (Pale) gene was targeted to assess the efficiency of the embryo transformations.

本综述探讨了昆虫性别决定通路（insect sex determination pathway）与CRISPR/Cas9基因编辑工具在基于遗传学的害虫防治方法中的应用。研究对新种Gonipterus sp. n. 2采用长读长与短读长测序技术开展全基因组从头测序（de novo genome），并结合RNA测序数据以及其他已发表组装基因组的鞘翅目物种的蛋白质序列完成了基因组组装与注释。本研究评估了该物种的基因家族扩张与收缩情况，重点聚焦于细胞色素P450（cytochrome P450）解毒基因；同时为基于显微注射的Gonipterus sp. n. 2胚胎CRISPR/Cas9转化实验方案奠定了研究基础，并以酪氨酸3-单加氧酶（Tyrosine 3-monooxygenase，简称Pale）基因为靶标，评估了胚胎转化实验的效率。