Self-limiting noctuids

A technology of Noctuidae and gender, applied in the field of splice boxes, can solve the problem of impractical physical separation

Pending Publication Date: 2021-01-05
OXITEC LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Large-scale rearing and physical separation of the sexes is impractical, or at least labor-intensive, in many cases

Method used

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Experimental program
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Effect test

Embodiment 1

[0223] The generation of embodiment 1.OX5403 fall armyworm

[0224] Plasmid pOX5403 ( figure 1 ) is based on the cloning vector pKC26-FB2 (Genbank #HQ998855). The plasmid backbone contains the pUC origin of replication and the β-lactamase gene that confers ampicillin resistance and is used in molecular cloning procedures. This plasmid portion is not contained in the rDNA or integrated into the insect genome.

[0225] pOX5403 also contains the complete rDNA incorporated into insects, including the synthetic DNA sequence encoding the DsRed2 red fluorescent marker protein (Clontech), the synthetic DNA sequence of the tetracycline-repressible transcriptional activator tTAV (based on the transcriptional activator from Escherichia coli and HSV-1 VP16 sequence fusion), and a modified Sfdsx splicing module derived from Spodoptera frugiperda. figure 2 The components shown in are detailed in Table 1. Plasmids were prepared by using conventional DNA cloning procedures.

[0226]The...

Embodiment 2

[0235] The generation of embodiment 2.OX5368 fall armyworm

[0236] Plasmid pOX5368 ( Figure 4 ) is based on the cloning vector pKC26-FB2 (Genbank #HQ998855). The plasmid backbone contains the pUC origin of replication and the β-lactamase gene that confers ampicillin resistance, which was used in molecular cloning procedures. This plasmid portion is not contained in the rDNA or integrated into the insect genome.

[0237] Plasmid pOX5368 also contains the complete rDNA incorporated into the insect, including the synthetic DNA sequence encoding the DsRed2 red fluorescent marker protein, the synthetic DNA sequence of the tetracycline repressible transcriptional activator tTAV2 (based on a fusion of sequences from the Escherichia coli and HSV-1 VP16 transcriptional activator) , with a modified Sfdsx splicing module derived from Spodoptera frugiperda. Figure 5 The components shown in are detailed in Table 2. Plasmids were prepared using conventional DNA cloning methods.

[0...

Embodiment 3

[0243] Example 3. Generation of OX5382 Spodoptera frugiperda

[0244] Plasmid pOX5382 ( Figure 7 ) based on the cloning vector pKC26-FB2 (Genbank #HQ998855). The plasmid backbone contains the pUC origin of replication and the β-lactamase gene that confers ampicillin resistance and can be used in molecular cloning procedures. This plasmid portion is not contained in the rDNA or integrated into the insect genome.

[0245] pOX5382 also contains the complete rDNA incorporated into insects, including the synthetic DNA sequence encoding the DsRed2 red fluorescent marker protein, the synthetic DNA sequence of the tetracycline-repressible transcriptional activator tTAV (based on the fusion of sequences from Escherichia coli and the HSV-1 VP16 transcriptional activator ), and a modified Sfdsx splicing module derived from Spodoptera frugiperda. Figure 8 The components shown are detailed in Table 3. Plasmids were prepared by Oxitec Ltd using conventional DNA cloning procedures.

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Abstract

The invention provides a Noctuid dsx splice cassette for expression of a gene of interest on a sex- specific basis, gene expression systems for imparting a self-limiting trait to transformed Noctuidae, as well as transgenic Noctuidae and methods of suppressing populations of Noctuidae and reducing, inhibiting or eliminating crop damage caused by the Noctuid insects.

Description

[0001] Cross References to Related Applications [0002] This application claims the benefit of U.S. Provisional Patent Application 62 / 649,912, filed March 29, 2018, which is incorporated herein by reference in its entirety. [0003] Sequence Listing Citation [0004] This application incorporates by reference the "Sequence Listing" (identified below) which is filed as a text file with this application. A text file copy of the Sequence Listing submitted with this article is labeled "Sequence Listing," is 131,925 bytes in size, and was created on March 27, 2019; the Sequence Listing is hereby incorporated by reference in its entirety. Background technique [0005] Noctuids, or the family Noctuidae, are known by a variety of common names, including cutworms, armyworms, and moths. Noctuidae covers more than 1,000 genera and more than 11,000 species. There are several genera and species within the Noctuidae family that cause massive crop damage and billions of dollars in losses...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A01K67/033C12N15/85C12N15/63
CPCA01K67/0339A01K2217/054A01K2227/706A01K2267/02C12N15/63C12N15/8509C12N2800/90C12N2830/42C12N2830/75C12N2840/44
Inventor 斯蒂芬·乔伊斯内森·罗斯凯利·马岑凯瑟琳·里夫露西·布鲁姆亚当·沃克西蒙·沃纳尼尔·莫里森
Owner OXITEC LTD
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