April

HP
During the campus open day, known as the Magnolia Festival, we showcased the charm of synthetic biology to students, teachers, and the public through a variety of creative formats, making the promotional activities engaging and entertaining.

May

Wet lab:
The experiments preliminarily validated that S.cerevisiae toxic genes MazF can be used.

HP
Created the initial draft of the Engineered Microorganism: Under the Sword of Damocles card game. Participated in the World Intelligence Conference (WIC) and introduced synthetic biology at our booth. Our theme covered the design and synthesis of artificial genomes, DNA storage, as well as the intelligent, automated, and flexible artificial cell factories.

June

Wet lab
S.cerevisiae：
pR_DK02、pR-DK03plasmid and control plasmid were separately transformed into BY4742 The experimental group showed minimal growth, while the control group grew normally, confirming that toxic genes can kill S.cerevisiae.

Test paper：
The experimental plan was initially determined.

HP
Improved and refined the card game rules.
Commenced a 14-day volunteering program, teaching students in rural areas about synthetic biology.

July

Wet lab:
Test paper:
Initial experiments attempted to replicate test papers according to literature. E.coli strains induced by IPTG to produce RFP were successfully immobilized on the test paper, but the desired luminescent effect was not achieved. Attempts to add LB medium to the test paper and take moisture-preserving measures successfully produced visible fluorescence after induction.

Kill Switch:
Constructed fusion proteins with dCas and Sir2.
Attempted to suppress the GFP gene using the constructed fusion protein and observed fluorescence intensity under a fluorescence microscope, confirming its inhibitory effect.
The plasmid containing the constructed fusion protein was co-transformed with pR-DK03 into BY4742. Compared to the control group, the strain grown showed a significant reduction, proving the effectiveness of EcoRI in killing the strain.
Conducted research on E.coli epigenetic modifications and preliminarily selected methylation as a method to suppress toxic genes.

HP：
Had our first dialogue with Tianjin High School, introducing the iGEM competition process, touring their laboratory, and listing the necessary equipment for experiments.
Completed the final version of the Engineered Microorganism: Under the Sword of Damocles card game and put it up for sale on Taobao.
During a high school summer camp, we taught high school students how to use microbial fluorescent proteins (RFP, GFP) for painting and provided them with hands-on laboratory experience.

Dry lab：
Through reviewing several research papers, we identified the models to establish and comprehensively considered the relevant factors associated with the models. Eventually, the toxic gene expression model was determined. Based on further in-depth research, we preliminarily constructed multiple dynamic models employing multivariate ordinary differential equations to solve the concentrations of toxic gene transcripts, the levels of epigenetic modifiers, the proportions of modified promoters, the cell death rate, and the expression levels of toxic genes. Literature was investigated to construct the first Cell Death Simulation Model.

August

Wet lab:
S.cerevisiae:
Construct pR_DK04 and pR_DK05 plasmids,respectively, and transform them into the BY4742. After verification, it was found that the transformed cells grew properly, indicating the suppression of the toxic genes.

E. coli:
Constructed pR_DK06 and pR_DK07 plasmids, transformed into E. coli, but sequencing yielded no results.
Re-extracted pR_DK06 and pR_DK07plasmids, and sequencing was successful.
Transformed these plasmids into MG1655 E. coli containing the endogenous Dam methyltransferase gene and JM110 E. coli lacking it. Observations showed that the pR_DK07 suppression effect was contrary to the expected target, so this promoter was eliminated.
Fluorescence intensity of pR_DK06 plasmids in both strains was measured using an enzyme-linked immunosorbent assay, preliminarily confirming the methylation suppression effect.
Attempted to construct pR_DK09 and pR_DK14 plasmids.

Test paper:
Tested the effect of different inducer concentrations on test paper brightness.

HP:
We co-hosted an online debate with the BIT-China team from Beijing Institute of Technology (BIT). The debate centered around "The Promotion/Hindrance of Scientific Research by the Formulation of Biosafety and Bioethics Regulations."
Had our second dialogue with Tianjin High School, where we taught high school students synthetic biology experiments and played card games together.

Dry Lab：
We have attempted to construct and solve equations for the expression levels of toxic genes and equations related to cell growth and death. The epigenetic modification model and promoter modification equation have been successfully established, laying the foundation for the preliminary form of the final model. In designing the promoter modification equation, we made efforts to consider promoter activity, and we have initially determined the corresponding differential equation forms. The steps for making a Cell Death Simulation Model were improved based on the wet lab experiment test paper.

September

Wet lab:
E. coli:
Sequencing results for pK_DK09 plasmid were correct.
The toxic gene ccdB segment was synthesized by a company.
The lab already had pR_DK12 plasmids transformed into E. coli.
Testing different sgRNAs for the suppression of toxic genes.
Constructing fusion proteins with dCas and Dam methyltransferase.
Constructed pR_DK10 plasmids.

Test Paper:
Tested luminescent effects of strains containing pR_DK12 plasmids immobilized on test paper.
Tested the influence of different inducer concentrations on fluorescence intensity.
Tested luminescent effects of strains containing pR_DK14 plasmids immobilized on test paper but abandoned further testing due to the absence of fluorescence.

HP：
Completed a promotional comic for community science outreach activities to raise awareness about potential biosafety issues in daily life.
Produced and published a comic about the history of synthetic biology for science outreach, which was released on our WeChat account.

Dry lab：
We have successfully designed and established the equation for toxic gene expression. Using computer modeling tools, we integrated the formulas into a application (App) interface, enabling the drawing of curves by dragging control values. Two model graphs were created: one depicting the interaction model between methyltransferase concentration (Met) and unmethylated modified promoter concentration (Pro) over time, and another describing the interaction model between protein content of toxic gene expression and cell count over time. The Cell Death Simulation Model has yielded the final results. Additionally, concise documentation for the experimental procedures has been composed in the Wiki section.

October

Wet lab：
Fluorescence verification of CRISPRi suppression effect.

Dry lab：
The Wiki section related to the dry lab experiments and the software section has been completed, with content optimization carried out for both sections.