(1) Team members' safety training

(2) Introduction of supervising teachers

from Professor Ming Chen

- Respect: mutual respect among team members.

- Responsibility: Emphasize the importance of taking responsibility for our actions.

- Risk: Foster a culture of innovation and encourage taking calculated risks.

- Resilience: Stress the need for strong resilience and the ability to bounce back from setbacks.

from Professor Fan Yang

Encourage us not to be burdened by past achievements but to boldly pursue innovation.

(1) Explanation of iGEM scoring criteria

(2) Sharing outstanding iGEM projects from previous years

(3) Sharing of previous projects by ZJU-China team

Conduct a brainstorming session based on the specific track

Conservation - Shiyi Lin

Track ideas:

- Plant disease monitoring

- Plant virus defense

- Plant-based recombinant vaccines

- Plant communication

- Plant microplastics

- Introduction of ethnobotany

- Endangered species detection

- Livestock disease treatment

- Antibiotic use in livestock farming

- Livestock vaccines

- Livestock mosquito control

Fundational Advance - Zhenning Yang

Track ideas:

- Modular Receptor Platform Development

- Orthogonal optogenetic systems

- Protein targeting for synthetic biology research

- Artificial organelles

- Optimization of chassis organisms or development of new chassis organisms

Other Inspiration:

- Bioproduction of melanin

- Weight loss interventions

Biomanufacture - Wenqing Ding

Track ideas:

- Fire-resistant wall coating

- Toilet cleaner

- Bacterial pets

- Non-damaging hair dye/permanent wave solution

- Safety airbags

Other Inspiration:

- Microbial production of meat substitutes

- Emergency biological drinking water

- E. coli modification and engineering

- Control of miRNA entry into exosomes through adapter modification RNA live cell in situ staining

- Drug addiction treatment (neurology)

Diagnostic - Linghao Xia

Track ideas:

- Vaginal infection test

- Trinucleotide repeat biosensor

- Visual bacterial biosensor

- RNA m6A sequencing

- Contrast agent

- Complementary peptides

- RNA sensor

Environment - Junjie Li

Track ideas:

- Pine wood nematode treatment

- Red fire ant pheromone

- Activated carbon and biological removal of methyl perchlorate

- Coral reef restoration

- Degradation of polycyclic aromatic hydrocarbons in asphalt roads

Therapeutics - Mingxuan Zhang

Track ideas:

- Brief overview of the RASER (Abnormal Signal Rewiring Effector Release) system and improvement directions

- Bacterial tryptophan reduction for immune suppression mitigation

- Orthogonal IL-2-controlled signal response

- Tooth decay treatment

Climate Crisis - Hanzhang Li

Track ideas:

- Main focus on Microbial Fuel Cells (MFC) for energy-saving and emission reduction

- Artificial cement

- Plastic recycling - transformation of a specific item

Conduct a brainstorming session based on the specific track

Shiyi Lin

- Brucellosis prevention and control

- Microplastic enrichment

- Bacterial imaging

- Bacterial neural networks

- E.coli with sound production function

- E.coli with hearing ability

- Bacterial long-distance signal transmission (quantum entanglement)

- Biopolymer films & bacteria color imaging & scroll repair

- Bacterial high-intensity thin films

Junjie Li

- Bacterial warfare/gene transfer

- Hangover cure

- Mosquito control

- Physical contact signaling

- Magnetic field-controlled engineering bacteriopharmaceutical delivery

- Antiviral bio-coatings

- Air purification wall coatings

- pH-regulating bacteria

Linghao Xia

- Yeast artificial organelles

- Modularization of photosensitive receptors

- Mirror information storage, conversion of chiral D to L

- Phase separation mechanism

Mingxuan Zhang

- RASER for the treatment of EGFR mutations

- Tumor hyperthermi

- Adenovirus treatment for hair loss

- Treatment of land desertification & salinization

- Tumor vaccines

Wenqing Ding

- Adenovirus treatment for hair loss

- Plant-derived animal antibodies

- Treatment of psoriasis

- Plant injection of animal antibodies

- Reprogramming of cancer cells

- Meningitis treatment

- Novel intestinal chip

- Drug delivery through the blood-brain barrier

- Optogenetics cage

- Endoplasmic reticulum stress

Qingyao Yan

- Tumor bomb

- Alcohol detoxification + alcoholic hepatitis

- Gut microbiota and thrombosis

- Artificial photoperiod in plants

- DNA barcodes to track food poisoning

- Electroactive microorganisms

- Smell robo

- DNA origami

- Bacterial sound collector

- Probiotic fermentation and transformation of traditional Chinese medicine

- Genetic code rewriting

- Bioadhesive for rapid hemostasis

Zhengning Yang

- Melanin synthesis

- T6SS

- DNA molecular encryption

- Microbial fertilizers for soil improvement

- Space synthetic biology

Hanzhang Li

- Hangover cure

- Hair loss

- New reporting methods

- Anti-aging

- Livestock disease monitoring

Yueran Shen

- Adenovirus treatment for hair loss

- In-body microplastics

- Biochips x synthetic biology

Summary of project design progress

- Brucella - phage therapy

- Introduction of animal antibodies-NLR protein in plants, vaccine delivery methods

- Blood-brain barrier delivery - improve RNA stability

- DNA encryption technology - signal-induced DNA inversion

- Space water cycle

- Biomineralization and dental caries treatment

- Bacterial timed bomb - time-controlled suicidal plasmid

(1) ZJU-China 2023 project topic voting

(2) Discuss project feasibility with mentors

The RNA vaccine technology for plants has received unanimous approval from the members of the Wet team. Space water cycle and biomineralization project design have become alternative projects.

(1) Report project topic to our PI.

(2) Discuss project feasibility with our PI.

[First Project - saRNA]

(1) Adding a signaling mechanism - Is it too burdensome for systemic delivery? For example, targeting rice blast disease in leaves No need for systemic delivery in space No need for continuous delivery over time

(2) Is NLR a mechanism in plants? What is the innovation point? Using antibodies produced by animals for enhanced specificity and resistance The antibodies we use are also derived from animals and their sequences can be traced

(3) How is it different from reference literature? The article still relies on transgenic methods (the principle of disease resistance remains unchanged, mainly focusing on implementation methods)

Issues: Complexity, need further clarification, feasibility needs to be thoroughly explored Improvement direction: Precision control rather than passive control, expressing at the right place and time (right place, right time to express and suicide - also applicable to the third project)

[Second Project - Mars]

(1) Only the initial story is related to Mars, the subsequent experiments are not strongly linked to Mars... Consider conditions of anoxic environment and extreme temperatures (suggestion: enhance the local environment, no need to react in an exposed environment)

(2) Are there no other aspects on Mars that need improvement besides modifying perchlorates?

(3) Based on this, further exploration can be done - determining water production

(4) (This implies creating artificial organisms that are entirely adapted to the Martian environment, which seems extremely challenging)

Improvement direction: Early investigation - Identify the remaining issues with soil composition Biosafety considerations for humans; Biosecurity mainly targeting the environment

[Third Project - Cartilage]

(1) Heart stent (rhythm heartbeat)

(2) Shaped cartilage, e.g., nose reconstruction, ear reconstruction

(3) Need to add new highlights (otherwise, it's a bit ordinary)

(4) Cartilage can be integrated using in-body 3D printing

(5) Cell 3D printing, I have heard reports where integrins are used to guide structure growth of cells

Improvement direction: Add shape control, benchmark current cartilage repair and treatment mechanisms

[Future Work]

Feasibility exploration: Technological roadmap, involved species, antigen-antibody expression system, biosafety concerns, environmental pollution (First project has higher risks but is more innovative; second and third projects have minimal risks)

Preliminary experiment design

(1) Self-replication-related sequences of plant positive-sense RNA viruses, such as Sindbis virus, TMV, BMV [by Wenqing and Junjie]

(2) Rice NLR/ Tobacco NLR/ Arabidopsis thaliana NLR: Further research and selection + specific sequence identification [by Qinyao and Mingxuan]

(3) VHH sequences: Fluorescent proteins, VHHs targeting pathogens [by Linghao and Hanzhang]

(4) TLS sequences [by Shiyi and Zhenni]

(5) Chassis research [by Yueran and Lu Qian]

(1) Summary of recent work

(2) Discussion of project issues

- PCR of pikm gene

- Primer design (3 types of TLS, Arabidopsis thaliana, Arabidopsis thaliana full, tobacco full)

- Gene PCR of TLS-GFP

- Homologous recombination

- Transformation into E.coli

- Internal innovation and entrepreneurship fund application

(1) Summary of recent work

(2) Discussion of project issues

- Agrobacterium-mediated transformation of TLS

- Alignment of pikm2 gene

- Preparation of Agrobacterium competent cells

- Primer design for cuticle protein

- Completion of multiplex PCR with multiple cloning sites

(1) Summary of recent work

(2) Discussion of project issues

- Went to Professor Liang Yan's place to collect the extracted cDNA

- Purchased tobacco seeds and growth medium

- Successfully conducted PCR amplification of the pikm2 gene

- Received primers for viral coat protein and fluorescent protein, ready for ligation

(1) Summary of recent work

(2) Discussion of project issues

- Construction of pikm2 plasmid

- Construction of 3 TLS plasmids

- Learning tobacco cultivation techniques

- Learning transient expression techniques

(1) Summary of recent work

(2) Discussion of project issues

- Software: This discussion mainly revolves around the functional prediction model based on amino acid sequence and structure, involving parameter learning, structure influence, chemical reactions, and discussing issues such as computational capabilities and structure file formats.

- Discussion on the issues of Agrobacterium, handling methods for variables, and testing results.

- Discussion on rice cultivation plan, progress of experiments, and next steps' arrangements.

(1) Summary of recent work

(2) Discussion of project issues

File handling, format confirmation, error debugging, and performance optimization in the process of model deployment and testing.

(1) Summary of recent work

(2) Discussion of project issues

- Summer work arrangement

- Preparation for the Conference of China iGEMer Community

- Presentation of the project design

- Discussing project feasibility with iGEM teams from other universities in China

(1) Summary of recent work

(2) Discussion of project issues

(3) Literature sharing

- Design and order the remaining 19 TLS DNA sequences for Arabidopsis thaliana

- Attempt to produce multiple copies of TLS, around a dozen

- The previous TMV clone recovery concentration was a bit low.

- The pgreen-pikm1 sample has been received

- Primers for ID sequence are ready; infect plants with TMV and use immunosuppressants

- Literature sharing by Zhenning

(1) Summary of recent work

(2) Discussion of project issues

(3) Literature sharing

- Agrobacterium activation completed

- Injected 4 sets of TLS into 16 tobacco plants

- Performed tRNA experiments on all 20 groups of Arabidopsis thaliana, connected them to GUS

- Completed ordering of repetitive tandem sequences

- Removed TMV coat protein+movement+pgreen for testing, only backbone without target gene, waiting for sequencing results for the remaining points

- Completed replacement of ID sequence for pikm1

- Designed a hardware cursor caliper puncher

- Changed the strategy of the model

- Software: Predicting fragments based on the chemical properties, hydrophobicity, and electric charge of exposed bases, real-time fragment simulation of compound/amino acid natural growth process, selecting method - computational efficiency, pure random process

- Literature sharing by Linghao and Yueran: Linghao - RDRP truncation; Yueran - downstream pathway under ETI

(1) Summary of recent work

(2) Discussion of project issues

- Injected TLS into tobacco plants, preparing to send 16 plants to the College of Agriculture to observe fluorescence.

- Human practice: Contacting the Food and Agriculture Organization of the United Nations and the Department of Agriculture and Rural Affairs of Zhejiang Province.

- Contacting the mentor.

- Failed to construct the rdrp plasmid.

- Modified the pikm1 sequence into nanobody; completed Agrobacterium-mediated transformation of pikm1, pikm2, and gfp.

- TMV plasmid construction.

- Brainstorming on the design of the biosafety module.

(1) Summary of recent work

(2) Discussion of project issues

- TLS fluorescence observation, qPCR preparation.

- Tobacco transient expression injection (with GFP stimulant), observe immune phenotype after 4-5 days.

- Software: Diffusion model used for protein structure prediction.

- Hardware: Puncher design completed.

- Idea for fluorescence tracking, capable of visualizing fluorescent movement.

(1) Summary of recent work

(2) Discussion of project issues

(3) Literature sharing

- Plant RNA extraction, negative control missing.

- Construction of TLS plasmid, already performed homologous recombination.

- Reconstructing the rdrp plasmid.

- Spottings phenotype, if not present, uses reactive oxygen species.

- Model: Infection model supplemented with invasion and invasion kinetics.

- Model: Considering not only the flow of RNA in the straight tube system, but also constructing a more realistic system.

- Literature sharing: Model of spore transmission and infection in the air.

(1) Summary of recent work

(2) Discussion of project issues

(3) Literature sharing

- The replicating tobacco plants for the four TLS groups have been prepared.

- Several new TLS constructs are not successful.

- Biosafety: Promoters, elements, terminators; validation of promoters and terminators.

- Software: Parameter tuning phase, running the model requires a server; software developed as a web tool.

- Hardware: Found a good approach, referring to a master's thesis from Zhejiang University.

- Literature sharing: Zhenning, Yueran.

(1) Summary of recent work

(2) Discussion of project issues

(3) Literature sharing

- New construction of TLS GUS

- Review of problems in rdrp plasmid construction

- Linearization of pgreen-tmv

- Activation and injection of pikm1-2

- New detection method for TLS

- Validation of promoter and terminator

- Enterprise contact for Human Practice; Discussion of project design with Professor Chai Jijie

- Consultation on legal regulations and related issues

- Contact with transformation author on WeChat public account

- Literature sharing: Ling Hao's paper on NLR modification; Qin Yao - the impact of aerosols on farmers (allergens)

(1) Summary of recent work

(2) Discussion of project issues

- TMV-GFP plasmid to be constructed, successful bacterial transformation, tobacco injection, immune phenotype; the previous weeks were preliminary experiments, phenotype expression will start this weekend.

- Verify TMV load by measuring GFP expression level, assess immune clearance; can observe photosynthetic changes and macroscopic changes.

- rdrp: new plasmid construction.

- TLS: 8 plasmids, transformed and plated for testing; different reporters, GFP, GUS, RUBY - plasmid construction and injection.

- Advanced modification: deletion of certain structures to investigate their impact on mobility.

- Professor Chen's suggestion: after obtaining results, combine them with dry experiments to predict which key structure affects RNA mobility.

- Construction of plasmids combining TLS and rdrp.

(1) Summary of recent work

(2) Discussion of project issues

- RUBY plasmid construction completed.

- GUS plasmid construction.

- GFP protein extraction finished.

- Tobacco injection for NLR, exploring concentration conditions.

- Software: data processing.

(1) Summary of recent work

(2) Discussion of project issues

- Hardware: Purchase plan and intentions to make a microscope and punching device

- The TLS experiment photos are still being processed.

- TLS mobility verification.

- Summary of wiki documentation and preparation for project closure.

Summary of recent work

Verification of the biosafety module and exploration of research using more toxic genes.

Conducting sunlight exposure experiments on the biosafety module, observing different time gradients, and placing 16 plates for observation.

The addition of local features with only single-point mutations has resulted in a decline in model performance, indicating that our data processing is still insufficient. I am currently contemplating implementing new advanced image processing methods through literature and other models, as well as exploring new approaches to handle this data.

Summary of recent work

It has been discovered that the design issue near the DMP protein has led to the deletion of the replication protein region while retaining this segment may potentially trigger an ATG initiation problem. Efforts are currently underway to resolve this issue.

By experimenting to expose the protein samples under sunlight, the lethal curve was observed. Currently, we are purchasing new toxic proteins and simultaneously attempting new promoters, but due to the inability to obtain inducers, the experiment has not yet begun.