SeBLOS2 knockout via CRISPR/Cas9 leads to the loss of larval integument coloration in Spodoptera exigua (Lepidoptera: Noctuidae).
Arch Insect Biochem Physiol
; 114(3): e22040, 2023 Nov.
Article
em En
| MEDLINE
| ID: mdl-37622407
ABSTRACT
CRISPR/Cas9 technology is a precise and powerful tool for functionally exploring insect genes. The present study tested CRISPR/Cas9 as a way of undertaking effective gene mutagenesis in an important agricultural pest, the beet armyworm Spodoptera exigua. Based on a S. exigua transcriptome database, the entire complementary DNA sequence of SeBLOS2 encoding 140 amino acid residues was cloned. The gene was highly expressed in late larval stages (L3-L5). Using the CRISPR/Cas9 method, SeBLOS2 was knocked out by altering two sites in the coding region. This resulted in 70%-74% of the G0 generation (L4-L5) larvae displaying mosaic translucent integument. Four different mutations occurred at SeBLOS2-specific target sites, as demonstrated by further polymerase chain reaction-based genotypic analysis. Homozygote mutant L3 larvae were obtained in the G1 generation, with complete loss of white stripes and spots on their larval integument. These results demonstrate a crucial role of SeBLOS2 in integument pigmentation and suggest that the gene can act as a suitable nonlethal marker for functional research on genes in S. exigua and other Lepidopteran pests.
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Base de dados:
MEDLINE
Assunto principal:
Tegumento Comum
/
Sistemas CRISPR-Cas
Limite:
Animals
Idioma:
En
Ano de publicação:
2023
Tipo de documento:
Article