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Dryad

Mutant Harmonia axyridis raw sequencing data of CRISPR/Cas9 induced mutations in the genes laccase2 and scarlet

Cite this dataset

Partosh, Tamir; Davidovitz, Michael; Firer, Noa; Pines, Gur (2024). Mutant Harmonia axyridis raw sequencing data of CRISPR/Cas9 induced mutations in the genes laccase2 and scarlet [Dataset]. Dryad. https://doi.org/10.5061/dryad.t1g1jwtb1

Abstract

Harmonia axyridis (Pallas), commonly known as the Asian lady beetle, is a native insect species of Asia that has been intentionally introduced to various regions for biocontrol purposes. However, its widespread presence beyond its original release sites suggests a high degree of invasiveness. In this study, we utilized the CRISPR-Cas9 approach to achieve precise genome editing in H. axyridis. Specifically, we targeted two genes in H. axyridis, laccase2 and scarlet, knockdown of which orthologs in other insects showed visible phenotypic changes. The knockout laccase2 resulted in an early-detectable phenotype but also in lethality. However, we successfully established a viable and genetically stable mutant colony by disrupting the scarlet gene, resulting in beetles with white eyes. Our findings contribute to the expanding knowledge of genetic manipulation in H. axyridis and provide insights into its potential for future research and practical applications for biocontrol and invasive species management.

README: Mutant Harmonia axyridis raw sequencing data of CRISPR/Cas9 induced mutations in the genes laccase2 and scarlet

https://doi.org/10.5061/dryad.t1g1jwtb1

In this study, we have edited two genes in the Asian Ladybeetle, Harmonia axyridis, using CRISPR/Cas9

The two genes are:

  • Laccase2 - This gene is responsible for generating melanin. Its knockout resulted in white patches throughout the larvae's bodies, but it proved lethal when widespread. Homozygosity could not be achieved.
  • Scarlet - This gene codes for a portion of a pigment pump in insects' eyes. The phenotype was seen only in the eyes of adult beetles; no physiological effects were seen, and a stable line was generated.

This dataset contains the sequence data as received from the sequencing provider. The sequences are divided into two groups. The first, "lac2," contains sequences obtained from H. axyridis with CRISPR-Cas9-induced mutation in the lac2 gene. The second group includes sequences obtained from mutants in the scarlet gene. 

Description of the data and file structure

The files are organized according to the figures, including supplementary. 

File names are composed of the target gene for editing and a running serial number

Each sequence has two file types: the chromatogram (trace) and a FASTA file representing the most likely sequence.

The chromatogram files are in a .ab1 format and can be opened using many different programs, including FinchTV: 

https://digitalworldbiology.com/FinchTV

Methods

This dataset includes the raw sequencing data of the mutated H. axyridis as provided by the sequencing vendor.