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Biology subjects

Lorenzen, M.

Publications and source records attributed to Lorenzen, M..

2 recordsLinked to original sources

Evaluation of Peregrinus maidis transformer-2 as a target for CRISPR-based control

The corn planthopper, Peregrinus maidis, is an economically important pest of corn and sorghum. Here we report the initial steps towards developing a CRISPR-based control method, precision guided sterile insect technique (pgSIT), for this hemipteran pest. Specifically, we evaluated the potential of transformer-2 (tra-2) as a target for sterilizing insects. First, we identified tra-2 transcripts within our P. maidis transcriptome database and performed RNA interference (RNAi) to confirm functional conservation. RNAi-mediated knockdown of Pmtra-2 in nymphs transformed females into pseudomales with deformed ovipositors resembling male claspers. While males showed no overt difference in appearance, they were indeed sterile. Importantly, the results were similar to those observed in another planthopper, Nilaparvata lugens. We also used CRISPR/Cas9 genome editing to assess the impact of tra-2 knockout in injectees. CRISPR-mediated knockout of Pmtra-2 had lethal effects on embryos, and hence not many injectees reached adulthood. However, mosaic knockout of Pmtra-2 did impact female and male fertility, which supports the use of tra-2 as a target for pgSIT in this hemipteran species.

genetics↗

Window to the Fluorescence: The White Eye-Color Gene of Western Corn Rootworm, Diabrotica virgifera virgifera

Eye-color mutations have proven useful in multiple insect species to help facilitate the development and use of transgenic tools for functional genomics. While there is species-specific variation in the pigments used to color insect eyes, every species studied thus far requires an ortholog of the ABC transporter gene white for proper pigmentation of the eyes. Previously, we generated transgenic western corn rootworm, Diabrotica virgifera virgifera, and found that their wild-type eye color obscured our ability to visualize a fluorescent marker driven by the widely used 3xP3 eye-specific promoter. Therefore, we sought to identify the D. v. virgifera ortholog of white (Dvvw). Here we report the discovery, cloning, and analysis of Dvvw cDNA and promoter. We also utilize RNA interference to knock down Dvvw mRNA in a transgenic strain, thereby demonstrating the utility of eye-color mutations when developing transgenic technologies.

genetics↗