PT - JOURNAL ARTICLE AU - Yuji Matsuoka AU - Taro Nakamura AU - Takahito Watanabe AU - Austen A. Barnett AU - Sumihare Noji AU - Taro Mito AU - Cassandra G. Extavour TI - Establishment of CRISPR/Cas9-based knock-in in a hemimetabolous insect: targeted gene tagging in the cricket <em>Gryllus bimaculatus</em> AID - 10.1101/2021.05.10.441399 DP - 2021 Jan 01 TA - bioRxiv PG - 2021.05.10.441399 4099 - http://biorxiv.org/content/early/2021/05/10/2021.05.10.441399.short 4100 - http://biorxiv.org/content/early/2021/05/10/2021.05.10.441399.full AB - Studies of traditional model organisms like the fruit fly Drosophila melanogaster have contributed immensely to our understanding of the genetic basis of developmental processes. However, the generalizability of these findings cannot be confirmed without functional genetic analyses in additional organisms. Direct genome editing using targeted nucleases has the potential to transform hitherto poorly-understood organisms into viable laboratory organisms for functional genetic study. To this end, here we present a method to induce targeted genome knock-out and knock-in of desired sequences in an insect that serves as an informative contrast to Drosophila, the cricket Gryllus bimaculatus. The efficiency of germ line transmission of induced mutations is comparable to that reported for other well-studied laboratory organisms, and knock-ins targeting introns yields viable, fertile animals in which knock-in events are directly detectable by visualization of a fluorescent marker in the expression pattern of the targeted gene. Combined with the recently assembled and annotated genome of this cricket, this knock-in/knock-out method increases the viability of G. bimaculatus as a tractable system for functional genetics in a basally branching insect.Competing Interest StatementThe authors have declared no competing interest.