An ovary-targeted nucleic acid delivery system, OT-NADPS, efficiently mediates the generation of transgenic silkworms.

Transgenic technology is crucial for advancing various areas of biological research. In silkworms, the traditional transgenic method relies on embryo microinjection, which is a labor-intensive process with limited efficiency and high technical barriers. To overcome these challenges, we developed an Ovary-Targeted Nucleic Acid Delivery Peptide, OT-NADPS, based on the Receptor-Mediated Ovary Transduction of Cargo system (ReMOT) by innovative integration. OT-NADPS, which consists of a nucleic acid loading domain and an ovarian targeting peptide, demonstrates strong nucleic acid binding capacity and excellent binding stability. It can efficiently deliver nucleic acid to silkworm ovaries through pupae injection, thereby enabling the generation of transgenic silkworms. Using the OT-NADPS delivery system, transgenic silkworms were obtained with a success rate of 3.92%, comparable to the 2.0%-5.0% success rate range in traditional embryo microinjection. This novel method shows a lower technical threshold and a higher success rate, which is potentially applied to transgenic operations in multiple insect species.