In this project, we are dedicated to sharing the software tools and component materials developed throughout our research journey. Beyond laboratory work, we have also actively engaged with communities, integrated synthetic biology education into public outreach, and promoted the concepts of sustainability.

In terms of component development, we took the CRISPR-Cas9 system as the technical core and the pJOE8999 plasmid as the vector backbone to successfully construct a dual-gRNA editing vector named pWLS. This vector can simultaneously express two gRNAs targeting the srf or itu gene clusters respectively, which significantly improves the knockout efficiency of large-fragment genes. Future iGEM teams can directly use this series of standardized components we constructed, eliminating the cost of repeated development.

In the aspect of experimental research, we clarified the regulatory roles of relA (positive regulation), cdaA (positive regulation) and cheB (negative regulation) genes in CLPs synthesis, proposed a regulatory model for CLPs synthesis, and screened out HMBY-106 (a mutant strain with strong antibacterial activity and high yield) as well as a more efficient relA-targeted mutant strain. These achievements not only provide key data for the research on the synthesis mechanism of CLPs, but also offer valuable insights and strategies for the industrial production and application of CLPs in the agricultural field in the future.

The software developed by the team serves as the core computational engine of this project, providing systematic acceleration for the strain optimization process. Through the rational design of COM domains and reprogramming of nonribosomal peptide synthetases (NRPS), the software significantly shortens the trial-and-error cycle of traditional experiments, enabling us to achieve quantifiable yield breakthroughs within a single research phase—with Fengycin production increased by 1.8-fold.

The precise predictive capability of the software platform concentrates experimental resources on the most promising genetic targets, which not only optimizes resource allocation efficiency but also greatly accelerates the overall research progress. As an open-source tool, this software transcends the scope of the project itself and delivers long-term reusable technical support to the synthetic biology community. Future iGEM teams can directly adopt the standardized workflow we established and apply it to various NRPS-related engineering projects, effectively lowering the project initiation threshold and time costs.

We have developed differentiated science popularization programs for diverse groups, including hearing-impaired children, kindergarteners, middle school students, college students, and international students, ensuring that every group has the opportunity to access and learn about synthetic biology, as well as experience the unique charm of iGEM (International Genetically Engineered Machine Competition).

Meanwhile, we have expanded the reach of our communication through multiple media channels: we reach a broad audience via radio stations and social media platforms, and deepen knowledge dissemination through offline interactive stalls and thematic lectures, comprehensively promoting the concepts of sustainable development. Furthermore, we have strengthened cooperation with universities, enterprises, and farmers, injecting more impetus into the sustainable advancement of the project through the university-industry-research collaboration model.