Genome-wide CRISPR screening in Spodoptera frugiperda reveals the determinants of insecticide activation and sensitivity. Nature Communications. The dataset comprises (i) raw RNA sequencing data from Sf9 cells, (ii) protein structure files of CYP333B enzymes used for molecular docking analyses and ligand structure files of six insecticides, (iii) amino acid sequences of CYP333B proteins, and (iv) Sanger sequencing data for validating gene editing. These data were generated in the context of a genome-wide CRISPR knockout screening system developed to investigate the molecular determinants of insecticide activation and sensitivity in Spodoptera frugiperda. The RNA-seq data enable transcriptomic analyses, while the protein and ligand structure files support structural and docking studies of cytochrome P450-mediated insecticide interactions. The protein sequences and Sanger sequencing further facilitate functional and gene editing analyses of CYP333B genes.

Dataset DOI: 10.5061/dryad.pk0p2nh43

Description of the data and file structure

Overview

This repository is associated with Wang et al. (2026). Genome-wide CRISPR screening in Spodoptera frugiperda reveals the determinants of insecticide activation and sensitivity. Nature Communications. In Press. The dataset includes RNA sequencing data, protein structure files used for molecular docking analyses, ligand structure files, protein sequences of CYP333B genes, and Sanger sequencing data supporting the study. Detailed descriptions of experimental procedures can be found in the associated publication.
Contents of the Dataset

File/Folder	Description
RNAseq_data	Contains raw RNA-seq data from Spodoptera frugiperda Sf9 cell line.
SF91_1.fq.gz	Paired-end RNA-seq read 1 for biological replicate 1.
SF91_2.fq.gz	Paired-end RNA-seq read 2 for biological replicate 1.
SF92_1.fq.gz	Paired-end RNA-seq read 1 for biological replicate 2.
SF92_2.fq.gz	Paired-end RNA-seq read 2 for biological replicate 2.
SF93_1.fq.gz	Paired-end RNA-seq read 1 for biological replicate 3.
SF93_2.fq.gz	Paired-end RNA-seq read 2 for biological replicate 3.
Docking_structures	Contains protein structure files and ligand files used for molecular docking analyses. These files can be opened and visualised using the Schrödinger Maestro Suite.
Sf9_CYP333B3_2.pdb	Predicted 3D structure of CYP333B3 from Sf9 cells.
Sf9_CYP333B4_0.pdb	Predicted 3D structure of CYP333B4 from Sf9 cells.
SfXZ_CYP333B3_4.pdb	Predicted 3D structure of CYP333B3 from SfXZ strain.
SfXZ_CYP333B4_1.pdb	Predicted 3D structure of CYP333B4 from SfXZ strain.
Chlorfenapyr.mol2	Ligand structure file of chlorfenapyr used for molecular docking analyses.
Diafenthiuron.mol2	Ligand structure file of diafenthiuron used for molecular docking analyses.
Fipronil.mol2	Ligand structure file of fipronil used for molecular docking analyses.
Phoxim.mol2	Ligand structure file of phoxim used for molecular docking analyses.
Pyridalyl.mol2	Ligand structure file of pyridalyl used for molecular docking analyses.
Tralomethrin.mol2	Ligand structure file of tralomethrin used for molecular docking analyses.
Protein_sequences	Contains CYP333B protein sequences.
Spodoptera_frugiperda_CYP333B_protein_sequences.fasta	FASTA file containing amino acid sequences of four CYP333B proteins. The files can be viewed and analysed using Geneious software.
Sanger_sequencing	Contains Sanger sequencing data used for validating gene editing.
Sanger_sequencing.zip	Raw Sanger sequencing files supporting gene editing. The contents of the compressed file are listed below. The file names follow the figure annotations in the associated manuscript. All files can be viewed and analysed using Geneious software.
Supplementary Fig. 1 E	Nucleotide sequences of indels induced by pB-CRISPR
Supplementary Fig. 1 F	Nucleotide sequences of indels induced by SfpB-CRISPR
Supplementary Fig. 12 C	Nucleotide sequences of indels induced by Sf9B4-g1
Supplementary Fig. 12 D	Nucleotide sequences of indels induced by Sf9B4-g2
Supplementary Fig. 12 E	Nucleotide sequences of indels induced by Sf9B4-g3
Supplementary Fig. 14	CRISPR-Cas9-mediated knockout of CYP333B4 in S. frugiperda (SfXZ-B4-KO strain)
Supplementary Fig. 15	CRISPR-Cas9-mediated knockout of CYP333B3 in S. frugiperda (SfXZ-B3-KO strain)
Supplementary Fig. 16	CRISPR-Cas9-mediated knockout of both CYP333B4 and CYP333B3 in S. frugiperda (SfXZ-B4B3-KO1 strain)
Supplementary Fig. 17	CRISPR-Cas9-mediated knockout of both CYP333B4 and CYP333B3 genes in S. frugiperda (SfXZ-B4B3-KO2 strain)

Data from: Genome-wide CRISPR screening in Spodoptera frugiperda reveals the determinants of insecticide activation and sensitivity

Data files

Abstract

Description of the data and file structure

Data from: Genome-wide CRISPR screening in Spodoptera frugiperda reveals the determinants of insecticide activation and sensitivity

Data files

Abstract

README: Data from: Genome-wide CRISPR screening in Spodoptera frugiperda reveals the determinants of insecticide activation and sensitivity

Description of the data and file structure