biostudies-literatureWang XNational Natural Science Foundation of Chinathe National Key R&D Program of China285https://www.ebi.ac.uk/biostudies/studies/S-EPMC12844060biostudies-literatureUnknown31(2)<i>Portunus trituberculatus</i> is an economically important marine crustacean in East Asia's aquaculture industry. Nevertheless, precise genome modification has not yet been established. In this study, we evaluated the applicability of the CRISPR-Cas9 gene editing system in <i>P. trituberculatus</i> using electroporation for efficient delivery of the Cas9-sgRNA complex into zygotes. We systematically investigated electroporation parameters, including buffer composition, voltage, capacitance, and pulse times. Our results showed that artificial seawater was a superior buffer to phosphate-buffered saline (PBS) and identified an effective electroporation condition of 600 V, 1 μF capacitance, and two pulses, resulting in approximately 72.7% fluorescent zygotes. Under these electroporated condiMolecules (Basel, Switzerland)Establishment of CRISPR-Cas9-Mediated Gene Editing in the Swimming Crab <i>Portunus trituberculatus</i>.PMC128440602022YFD240010432341058 and 31972783Liu LShi CWang CWang XChen XLi RZhou YSong WZhao YMu CfalseEstablishment of CRISPR-Cas9-Mediated Gene Editing in the Swimming Crab <i>Portunus trituberculatus</i>.<i>Portunus trituberculatus</i> is an economically important marine crustacean in East Asia's aquaculture industry. Nevertheless, precise genome modification has not yet been established. In this study, we evaluated the applicability of the CRISPR-Cas9 gene editing system in <i>P. trituberculatus</i> using electroporation for efficient delivery of the Cas9-sgRNA complex into zygotes. We systematically investigated electroporation parameters, including buffer composition, voltage, capacitance, and pulse times. Our results showed that artificial seawater was a superior buffer to phosphate-buffered saline (PBS) and identified an effective electroporation condition of 600 V, 1 μF capacitance, and two pulses, resulting in approximately 72.7% fluorescent zygotes. Under these electroporated condi2026-01-01T00:00:00Z2026 Jan2026-06-10T03:17:55.591Z2026-06-10T03:12:52.2ZS-EPMC128440604159933410.3390/molecules31020285