CRISPR mutagenesis of Tsp29Fa and Tsp29Fb. (A) Genomic region encoding Tsp29Fa and Tsp29Fb on chromosome 2L (green line). Blue arrows indicate span and direction of Tsp29Fa and Tsp29Fb genes. Each gene has three predicted transcripts (denoted RA, RB, and RC), where beige boxes represent noncoding and purple boxes represent coding regions of exons (adapted from Flybase, release 6.20). Red arrows point to locations where guide RNAs gRNA1 and gRNA2 were targeted for CRISPR-Cas9 gene editing. (B) Predicted amino acid sequences encoded by Tsp29^J1a (left) and Df(2L)L7a (right) deletions. Green indicates unaltered Tsp29Fa amino acid sequences and purple indicates unaltered Tsp29Fb amino acid sequence. *, stop codon. Red underline marks transmembrane domain. (C) ClustalW alignment of mouse CD63 with Drosophila Tsp29Fa and Tsp29FB. Red boxes indicate the four conserved transmembrane domains, based on predictions from UniprotKB for mouse CD63. (D) Bar graph showing qRT-PCR analysis of Tsp29Fb mRNA levels in control and Tsp29Fa^J1a L3 larvae normalized to Δ-tubulin84B mRNA. n = 3 independent experiments. Error bars indicate standard deviation.