DIPA-CRISPR is a simple and accessible method for insect gene editing

Current approaches for insect gene editing require microinjection of materials into early embryos. This severely limits the application of gene editing to a great number of insect species, especially to those whose reproduction systems preclude access to early embryos for injection. To overcome these limitations, we report a simple and accessible method for insect gene editing, termed “direct parental” CRISPR (DIPA-CRISPR). We show that injection of Cas9 ribonucleoproteins (RNPs) into the haemocoel of adult females efficiently introduces heritable mutations in developing oocytes. Importantly, commercially available standard Cas9 protein can be directly used for DIPA-CRISPR, which makes this approach highly practical and feasible. DIPA-CRISPR enables highly efficient gene editing in the cockroaches, on which conventional approaches cannot be applied, and in the model beetle Tribolium castaneum. Due to its simplicity and accessibility, DIPA-CRISPR will greatly extend the application of gene editing technology to a wide variety of insects. [Display omitted] •A simple and efficient method for insect gene editing•Based on direct adult injection of Cas9 ribonucleoproteins•Readily implementable in non-specialist laboratories•Applicable to a wide diversity of non-model insects With over a million species described, insects are a treasure trove of diversity and represent boundless possibilities as research tools for answering fundamental questions in biology. Current approaches for insect gene editing require microinjection of materials into early embryos, which is highly challenging in most species. In this work, we established and optimized a simple and efficient method for insect gene editing by adult injection, which can be readily implemented in any laboratory and directly applied to a great diversity of non-model insect species. Shirai et al. develop a simple and accessible method, DIPA-CRISPR, for insect gene editing by direct adult injection of Cas9 ribonucleoproteins. Using it, they successfully establish gene knockouts in cockroaches, in which conventional embryo microinjection cannot be applied. Given its simplicity and versatility, DIPA-CRISPR has the potential to greatly extend the application of gene editing technology to a wide diversity of insects.