We found a match
Your institution may have access to this item. Find your institution then sign in to continue.
- Title
CRISPR/Cas9-Mediated Mutagenesis of Antennapedia in Spodoptera frugiperda.
- Authors
Wang, Congke; Zhao, Te; Liu, Xiaolong; Li, Tianliang; He, Leiming; Wang, Qinqin; Wang, Li; Zhou, Lin
- Abstract
Simple Summary: Spodoptera frugiperda, a significant pest affecting various crops, has been the focus of study to understand the role of Antennapedia (Antp), essential for insect thorax and wing development. We investigated Antp in S. frugiperda using RT-qPCR and CRISPR/Cas9 genome editing. We found that Antp is highly similar across Lepidoptera and is expressed throughout the life cycle of S. frugiperda, with the highest expression in the egg stage and significant expression between 12 to 48 h. The gene was primarily active in the thorax and legs. By knocking out Antp by CRISPR/Cas9, we observed abnormal development of the thoracic legs of the larvae and abnormal pupation. After the expression of Antp decreased, the expression of other Hox genes, appendage development genes and cuticular protein genes decreased significantly. The homeotic gene Antennapedia (Antp) has been identified as playing a pivotal role in the morphogenesis of the thorax and wings across various insect species. Leveraging insights from previous studies, the functional characterization of Antp in S. frugiperda was undertaken using RT-qPCR and the CRISPR/Cas9 genome-editing system. Phylogenetic analyses indicate that Antp shares a high degree of sequence homology among Lepidoptera species. The expression profile of SfAntp was detected by RT-qPCR. The results showed that SfAntp was expressed in the whole growth cycle of S. frugiperda, the expression level was the highest in the egg stage, and the expression level was higher from 12 h to 48 h. Tissue-specific expression profiling demonstrated that SfAntp was most abundantly expressed in the thoracic segments and legs. To functionally disrupt SfAntp, two sgRNA sites were designed at the first exon of SfAntp and the gene was knocked out by CRISPR/Cas9 via microinjection. The results showed that the deletion of SfAntp produced a mutant phenotype of thoracic fusion, thoracic leg defect, leg-like protrusions between the head and thoracic segments and pupation deformity. In addition, deletion of SfAntp resulted in high embryo mortality. Through DNA sequencing, it was found that the target site of the SfAntp mutant had different degrees of frameshift mutations, indicating that the mutant phenotype was indeed caused by the knockout of SfAntp.
- Subjects
FALL armyworm; HOMEOBOX genes; CRISPRS; LIFE cycles (Biology); MUTAGENESIS
- Publication
Insects (2075-4450), 2024, Vol 15, Issue 1, p16
- ISSN
2075-4450
- Publication type
Article
- DOI
10.3390/insects15010016