Targeting Sex Determination to Suppress Mosquito Populations

We previously developed a precision-guided Sterile Insect Technique (pgSIT) approach, which leveraged the CRISPR-Cas9 technology to simultaneously disrupt two genes essential for female viability and male fertility during the development and generate sex-sorted and sterilized males. In this work, we show that CRISPR-Cas9 can be used to simultaneously disrupt three genes in Ae. aegypti, dsx, intersex (ix), and beta-Tubulin 85D (betaTub), resulting in female lethality or masculinization and nearly complete male sterility.