CRISPR/Cas9-based split homing gene drive targeting doublesex for population suppression of the global fruit pest Drosophila suzukii.
CRISPR/Cas9-based split homing gene drive targeting doublesex for population suppression of the global fruit pest Drosophila suzukii.
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DOI：
10.1073/pnas.2301525120
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发表时间：
2023-06-20
影响因子：
11.1
通讯作者：
Scott, Maxwell J.
中科院分区：
文献类型：
--
作者：
Yadav, Amarish K.;Butler, Cole;Yamamoto, Akihiko;Patil, Anandrao A.;Lloyd, Alun L.;Scott, Maxwell J.
斑翅果蝇（Drosophila suzukii）是一种醋蝇，在世界范围内分布，对软皮水果造成重大经济损失。我们制作并评估了分裂CRISPR归巢基因驱动器，该驱动器靶向doubletex基因的保守雌性特异性外显子，该外显子对果蝇的性发育至关重要。我们的研究结果表明，归巢基因驱动器可以提供一个具有成本效益的方法来抑制D。铃木种群基于遗传的方法为控制害虫提供了环境友好的物种特异性方法。一种方法是CRISPR归巢基因驱动，靶向发育必需的基因，可以提供非常有效且具有成本效益的控制。虽然在开发蚊子疾病载体的归巢基因驱动方面取得了重大进展，但在农业害虫方面进展甚微。在这里，我们报告的分裂归巢驱动器的发展和评估，目标doublesex（dsx）基因的果蝇Suzukii，一种入侵害虫的软皮水果。由dsx单向导RNA和DsRed基因组成的驱动组件被引入dsx的女性特异性外显子中，这对于女性而不是男性的功能是必不可少的。然而，在大多数菌株中，半合子雌性不育并产生雄性dsx转录本。通过包括最佳剪接受体位点的修改的归巢驱动，来自四个独立系中的每一个的半合子雌性是可育的。用表达Cas9的细胞系观察到DsRed基因的高传递率（94至99%），Cas9具有来自D.铃木纳米启动子。在Cas9切割位点附近具有小的框内缺失的dsx的突变等位基因没有功能，因此不会提供对驱动的抗性。最后，数学模型表明，菌株可以用于抑制实验室笼种群的D。Suzukii以相对低的释放比（1：4）重复释放。我们的结果表明，分离的CRISPR归巢基因驱动菌株可能为控制D.铃木种群
Spotted wing drosophila (Drosophila suzukii) is a vinegar fly with a worldwide distribution that does significant economic damage to soft-skinned fruits. We made and evaluated split CRISPR homing gene drives that target the conserved female-specific exon of the doublesex gene, which is essential for sexual development in Drosophila. Our results suggest that homing gene drives could provide a cost-effective approach for suppression of D. suzukii populations. Genetic-based methods offer environmentally friendly species-specific approaches for control of insect pests. One method, CRISPR homing gene drive that target genes essential for development, could provide very efficient and cost-effective control. While significant progress has been made in developing homing gene drives for mosquito disease vectors, little progress has been made with agricultural insect pests. Here, we report the development and evaluation of split homing drives that target the doublesex (dsx) gene in Drosophila suzukii, an invasive pest of soft-skinned fruits. The drive component, consisting of dsx single guide RNA and DsRed genes, was introduced into the female-specific exon of dsx, which is essential for function in females but not males. However, in most strains, hemizygous females were sterile and produced the male dsx transcript. With a modified homing drive that included an optimal splice acceptor site, hemizygous females from each of the four independent lines were fertile. High transmission rates of the DsRed gene (94 to 99%) were observed with a line that expressed Cas9 with two nuclear localization sequences from the D. suzukii nanos promoter. Mutant alleles of dsx with small in-frame deletions near the Cas9 cut site were not functional and thus would not provide resistance to drive. Finally, mathematical modeling showed that the strains could be used for suppression of lab cage populations of D. suzukii with repeated releases at relatively low release ratios (1:4). Our results indicate that the split CRISPR homing gene drive strains could potentially provide an effective means for control of D. suzukii populations.
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DOI：
10.1079/9781800621176.0009
发表时间：
2022-01-01
期刊：
TRANSGENIC INSECTS, 2 EDITION
影响因子：
--
作者：
Champer, Jackson
通讯作者：
Champer, Jackson
影响因子：
5.4
作者：
Concha C;Palavesam A;Guerrero FD;Sagel A;Li F;Osborne JA;Hernandez Y;Pardo T;Quintero G;Vasquez M;Keller GP;Phillips PL;Welch JB;McMillan WO;Skoda SR;Scott MJ
通讯作者：
Scott MJ
影响因子：
5.4
作者：
Ant T;Koukidou M;Rempoulakis P;Gong HF;Economopoulos A;Vontas J;Alphey L
通讯作者：
Alphey L
影响因子：
7.7
作者：
Baeza-Centurion P;Miñana B;Valcárcel J;Lehner B
通讯作者：
Lehner B
DOI：
10.1073/pnas.1720354115
发表时间：
2018-05-22
影响因子：
11.1
作者：
Champer, Jackson;Liu, Jingxian;Messer, Philipp W.
通讯作者：
Messer, Philipp W.