Method for constructing anti-spinetoram neoseiulus californicus and ovarian targeting peptide thereof

By successfully knocking out the NcnAChRα6 gene in *Neoseiu Seymite* California using ReMOT Control technology, an ethyl spinosad-resistant strain with 23-fold resistance was established. This solved the problem of difficulty in improving drug resistance in *Neoseiu Seymite* California using existing technologies, and achieved stable genetics and efficient strain construction.

CN121406646APending Publication Date: 2026-01-27NANJING AGRICULTURAL UNIVERSITY
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Patent Information

Application Number
CN202511285766.3
Authority / Receiving Office
CN · China
Patent Type
Applications(China)
Current Assignee / Owner
Filing Date
2025-09-10
Publication Date
2026-01-27

AI Technical Summary

Technical Problem

Existing technologies make it difficult to efficiently improve drug resistance in Neoseido Seymite California through embryo microinjection, especially by knocking out drug resistance receptor genes. Moreover, the process is laborious and it is difficult to achieve stable genetic improvement of homozygous strains.

Method used

Using ReMOT Control technology, the ovarian-targeting peptide NcvgP2C and the Cas9 fusion protein NcvgP2C-Cas9, combined with sgRNA and saponins, were directly injected into the hemolymph of female adult Neoseis caecilians to edit the NcnAChRα6 gene. Subsequently, the strain was purified to obtain ethyl spinosad resistant strains.

Benefits of technology

A homozygous spinosad-resistant strain with 23 times the resistance was successfully established in Neoseido Seymite of California, reducing time and cost, expanding the possibilities for gene function research, and improving the compatibility of biological control.

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Abstract

The invention discloses a method for constructing an anti-spinetoram neoseiulus californicus gene and an ovary targeting peptide of the anti-spinetoram neoseiulus californicus gene. According to the invention, a segment of yolk protein targeted peptide binding region is identified in Neoseiulus californicus, and the yolk protein targeted peptide binding region and a Cas9 sequence are fused and constructed on a PET28a carrier, and the NcvgP2C-Cas9 protein is obtained through prokaryotic expression. NcvgP2C-Cas9 protein, sgNcnAChR alpha 6 and an endosome escape reagent are mixed and injected into female adult mites, NcnAChR alpha 6 gene large fragment deleted neoseiulus californicus offspring is obtained and shows a phenotype with 23.2 times of resistance to spinetoram, and the resistance of a knockout strain to the spinetoram belongs to an autosome partial recessive genetic mode. The invention provides a new method for the genetic improvement of the drug resistance of Neoseiulus californicus, and enhances the compatibility between biological control and pesticide treatment strategies.
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