mini embryo collection cage  (Genesee Scientific)

Journal: Nature Communications

Article Title: A temperature-sensitive CRISPR-Cas12a system for sterile insect technique

doi: 10.1038/s41467-025-64685-4

Figure Lengend Snippet: a We targeted the following loci: (1) βtubulin ( βtub ) which is testis-specific and causes male sterility when mutated, was targeted by four gRNAs (g1–g4); (2) sex lethal ( sxl ), which causes female lethality when disrupted, was targeted by four gRNAs (g1–g4); (3) transformer ( tra ), where targeting the female-specific exon results in intersex individuals was targeted by two gRNAs; and (4) double sex ( dsx ), where targeting the female-specific transcript leads to intersex phenotypes was targeted by one gRNA. Boxes indicate the exons and specific female and male transcripts are shown in different colors. Red vertical lines indicate premature stop codons. b To generate transgenic insects, gRNA-expressing constructs were built with mini-white, CFP/GFP markers for selection and tracking, and tRNA-based gRNA expression under the Drosophila U6-3 promoter. Constructs were integrated into the genome using PhiC31-mediated recombination.

Article Snippet: We employed 60 × 15 mm grape plates that were ordered from Lab Express (Cat. # 7007-60), and Drosophila Embryo Collection Cages from Genesee Scientific (Cat. # 59-100).

Techniques: Sterility, Transgenic Assay, Expressing, Construct, Selection

Journal: Nature Communications

Article Title: A temperature-sensitive CRISPR-Cas12a system for sterile insect technique

doi: 10.1038/s41467-025-64685-4

Figure Lengend Snippet: a To generate transgenic insects, gRNA-expressing constructs were built with mini-white, CFP/GFP markers for selection and tracking, and tRNA-based gRNA expression under the Drosophila U6-3 promoter. Constructs were integrated into the genome using PhiC31-mediated recombination. b Females carrying Cas12a were crossed to homozygous males carrying gRNAs targeting the βtubulin gene (G0). G1 males carrying both transgenes were then single pair-crossed to wildtype females to evaluate male sterility. As shown in the G2 table, these experiments were performed at 18 °C ( n = 8) and at 29 °C ( n = 15). c Editing rates at the two different target sites are shown for the βtub gene at 18 °C and 29 °C. For both temperatures, target sites were evaluated using two biological replicates or independent samples, each replicate containing 10 flies. Data are presented as mean values +/− SD. d Cas12a females were crossed to heterozygous males carrying gRNAs targeting sex lethal gene (G0). Then, G1 progeny was analyzed after one generation (G1). As shown in the G1 tables, these experiments were performed at 18 °C ( n = 15) and at 29 °C ( n = 17); average percentages from all experiments are shown in each table for the genotypes observed. e Editing rates at the two different target sites are shown for the sxl gene at 18 °C and 29 °C. For both temperatures, target sites were evaluated using two biological replicates or independent samples, each replicate containing 10 flies. Data are presented as mean values +/− SD.

Article Snippet: We employed 60 × 15 mm grape plates that were ordered from Lab Express (Cat. # 7007-60), and Drosophila Embryo Collection Cages from Genesee Scientific (Cat. # 59-100).

Techniques: Transgenic Assay, Expressing, Construct, Selection, Sterility