VGV-1

Although gene-editing breeding demonstrates significant potential in aquaculture, the field still lacks efficient gene delivery systems. Fish and crustacean embryos typically possess hard egg membranes or thick shells, making conventional microinjection inefficient and causing high mortality rates. As a result, there is an urgent need for a universal, highly efficient, and low-toxicity noninvasive delivery platform. In this study, we investigated the potential of high-efficiency, large-scale transfection using nanocarrier technology, renowned for its biocompatibility and efficient molecular payload encapsulation. Our research focused on developing a novel nanocarrier system, termed TNP, composed of a poly(ethylene glycol)-poly(lactide-co-glycolide) block copolymer (PEG-b-PLGA) and a cell-penetrating peptide (TAT, transactivator of transcription). The TNP nanocarrier enabled noninvasive delivery of genetic cargo, including eGFP-mRNA and CRISPR/Cas9 components, achieving high-throughput transfection in both shrimp and zebrafish embryos with successful transgene expression and targeted gene editing. We demonstrated that TNP efficiently traversed embryonic barriers, leveraging to facilitate high-payload gene delivery while maintaining minimal cytotoxicity and high embryo viability based on the transmembrane functionality of TAT peptide. These findings highlight the TNP has the potential to transform aquaculture genetic engineering, enabling trait enhancement such as disease resistance and accelerated growth. Furthermore, this study establishes a versatile and efficient gene-editing platform for aquatic species, addressing the critical demand for sustainable and ethically responsible biotechnological innovations in global seafood production.

It is of great significance for the discrimination of 2,4,6-trinitrotoluene (TNT) and 2,4,6-trinitrophenol (TNP) from their analogues. Here, based on the electron-deficient properties of TNT and TNP, a series of o-phenylenediamine/polyethyleneimine carbonized polymer dots (OPD/PEI CPDs) with different densities and types of amine groups (-NH₂) on the surface were designed by modulating the ratio of the precursors OPD and PEI. The surface -NH₂ group of OPD/PEI CPDs could form a Meisenheimer complex with TNT, triggering the Forster resonance energy transfer, while forming hydrogen bonds with hydroxyl in TNP, triggering the charge transfer and spectral overlap to produce photoinduced electron transfer accompanied with inner filter effect. When the ratio of OPD to PEI is 1:1 and the surface -NH₂ content is 26.06 %, the OPD/PEI₁ CPDs demonstrated a more superior sensing performance toward targets, including a low limit of detection (TNT: 324 nM and 255 µM, TNP: 21.08 nM and 318.6 nM), a rapid response (<1 s), and a rather good selectivity in the presence of 20 interferents. Moreover, the practicality of the OPD/PEI₁ CPDs was further verified by an OPD/PEI₁ CPDs-based paper sensor, which is capable of discriminating TNT and TNP particles and vapors as low as pg-level and ppm-level respectively.