title: Genome Editing ofWnt-1, a Gene Associated with Segmentation, via CRISPR/Cas9 in the Pine Caterpillar Moth,Dendrolimus punctatus.
abstract: The pine caterpillar moth,Dendrolimus punctatus, is a devastating forest pest. Genetic manipulation of this insect pest is limited due to the lack of genomic and functional genomic toolsets. Recently, CRISPR/Cas9 technology has been demonstrated to be a promising approach to modify the genome. To investigate gene functions during the embryogenesis, we introduced CRISPR/Cas9 system inD. punctatusto precisely and effectively manipulate gene expressions inmutant embryos. Compared to controls, knocking out ofDpWnt-1, a gene well known for its role in the early body planning, led to high embryonic mortality. Among these mutants, 32.9% of the embryos and larvae showed an abnormal development.DpWnt-1mutants predominantly exhibited abnormal posterior segments. In addition, multiple phenotypes were observed, including the loss of limbs and the head deformation, suggesting thatDpWnt-1signaling pathway is necessary for anterior segmentation and appendage development. Overall, our results demonstrate that CRISPR/Cas9 system is feasible and efficient in inducing mutations at a specific locus inD. punctatus. This study not only lays the foundation for characterizing gene functions in a non-model species, but also facilitates the future development of pest control alternatives for a major defoliator.
authors.affiliation: Key Laboratory of Forest Protection, Research Institute of Forest Ecology, Environment and Protection, Chinese Academy of Forestry, State Forestry Administration Beijing, China.
authors.forename: Huihui
authors.initials: H
authors.lastname: Liu
authors.affiliation: Key Laboratory of Insect Developmental and Evolutionary Biology, Institute of Plant Physiology and Ecology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences Shanghai, China.
authors.forename: Qun
authors.initials: Q
authors.lastname: Liu
authors.affiliation: Department of Entomology, University of Kentucky Lexington, KY, USA.
authors.forename: Xuguo
authors.initials: X
authors.lastname: Zhou
authors.affiliation: Key Laboratory of Insect Developmental and Evolutionary Biology, Institute of Plant Physiology and Ecology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences Shanghai, China.
authors.forename: Yongping
authors.initials: Y
authors.lastname: Huang
authors.affiliation: Key Laboratory of Forest Protection, Research Institute of Forest Ecology, Environment and Protection, Chinese Academy of Forestry, State Forestry Administration Beijing, China.
authors.forename: Zhen
authors.initials: Z
authors.lastname: Zhang
keywords: CRISPR/Cas9; Dendrolimus punctatus; Wnt-1; embryogenesis; genome editing; segmentation