Organization: Beijing Normal University

position:Professor

Why do we talk to him?

Professor Wang Zhanxin is an expert in molecular biology and holds a professorship at Beijing Normal University.

Our project was in the final stage of design, but we have encountered some obstacles in the specific design of slinker protein junction. Therefore, we hope to find experts in molecular biology to express their opinions and provide some suggestions on our current design, so as to help us overcome the difficulties and complete the final design.

Professor Wang Zhanxin is an expert in molecular biology and holds a professorship at Beijing Normal University.

our takeaways:

1.In terms of project feasibility:

I.Professor Wang Zhanxin emphasized that the stable connection between SBP and silica is crucial for the success of this project. Despite the non-covalent nature of the SBP protein and silica bond, the KD value indicates a strong connection between SBP and silica.

II. Professor Wang suggested in the design phase that we should pay more attention to whether this system is compatible with the pH levels in the human body.

2.Regarding the design:

In the initial design phase, our goal is to employ machine learning to enable bacteria to autonomously select the most suitable linker length, thereby overcoming potential steric hindrance effects during the connection process. Professor Wang Zhanxin noted that the length of the linker may not be the primary focus. As long as our linker is sufficiently long, it should be capable of achieving the connection, given that our screening objective is simply to choose the most appropriate length.

On June 30, 2023, with the support of the School of Life Sciences at Beijing Normal University, the Beijing iGEMer Exchange Conference organized by the BNU-China team was successfully held in Room 101 of Teaching Building 3 at Beijing Normal University. The conference brought together a total of over fifty participants from five teams: BNU-China, BUCT, BUCT-China, CAU-China, and Peking.

Each team presented their projects, and after discussing the content, they also provided suggestions and exchanged ideas with one another. Everyone spoke freely, engaging in a back-and-forth exchange of thoughts with different teams, benefiting from the collision of ideas and learning from each other's experiences.

Organization: Shenzhen Xinsu Technology

Why do we talk to him?

Dong Yiming, as the Partner and Chief Scientist of SiBio, is actively involved in interdisciplinary research in biology and other fields. We are seeking her advice on the industrialization aspects of our project.

She has previously participated in the iGEM competition and successfully guided a high school team to win the finalist. Due to recent changes in the iGEM rules, we would appreciate some advice regarding award applications for the competition this year.

our takeaways:

1.In terms of industrialization, she recommended intensifying communication with biotech companies to conduct a thorough analysis of the advantages our project holds over traditional modification methods. This strategic approach will grant us valuable insights into both the present market landscape and future market trends.

For the upstream, it is strongly advised to establish contact with companies specialized intraditional nano-silica modification. Engaging with them will allow us to inquire about the challenges they have encountered in this particular domain. Alternatively, reaching out to nanomaterial companies can help us explore their familiarity with the wide-ranging applications of modified nano-silica.

For the downstream, try to connectwith customers who are already utilizing modified nano-silica in their applications is crucial. This direct engagement will yield valuable feedback on the challenges they encounter. Additionally, it would be advantageous to approach companies specializing in gene and drug delivery to conduct a comprehensive analysis of the disparities, advantages, and disadvantages between various delivery methods and nano-silica delivery.

2.Regarding the iGEM competition, she recognized the innovative nature of our project and strongly suggested that we consider submitting it for awards within the "part" category.

Organization: Beijing Normal University

Position:Professor

Why do we talk to him?

To vertify the functions of our silinker,for example,to see if Twisted Silinker(TS) and Paring Silinker(PS) change in accordance with our idea after dimerization binding,to measure the value of KD and so on,we interviewed Professor Junjie Zhang from Beijing Normal University, an expert in biochemistry and molecular biodynamics.We hope to get more detailed information about protein binding and some professional suggestions on involved proteins of our project.

Our takeaways

1.about SBP : Junjie Zhang suggested us trying “the selected best SBP” provided in the paper first. If the SBP doesn’t work,then design and express our own polypeptides,and do the elution to select eligible SBP.This provides a feasible idea for our follow-up experiments.Meanwhile,he pressed that we shouldn’t add redundancy parts since they can bring pressure to our linker binding.We pay extra attention to this when designing.

2.about Paring Slinker:Junjie Zhang said detecting fluorescent signal is a good choice for us to vertify if proteins get dimerization,which is exactly what we were thinking.Also,he mentioned CytC has a function of oligomerizing proteins.We wonder if we can optimize our detection mode by releasing and binding CytC.

3.about Twisted Silinker：The TS we designed involves the change of proteinopathy,which is hard to observe.So we need to speculate about it by the change of fluorescent signal.Combining with the appliment of TS——drug delivery,Junjie Zhang told us the recent experiments of drug delivery often use flurescent protein to reflect durg encapsulation.But he also raised two questions:

I.saturation paradox：On the one hand,TS can’t be too dense while loading drugs.On the other hand,TS should be dense to avoid drug leak.

II.Reversibility：The command of loading and releasing drugs requires reversibility of the proteinopathy’s change.

4、about the appliment：Junjie Zhang thought as our system works extracellularly,we can shift our focus from calcium to other extracellular signals,such as heavy-metal ion,biotoxins.And we can even try to apply it to blood test and such more kinds of IVD.This points out a bright prospect of our project,and thus there’re more potential stakeholders.

Professor Junjie Zhang gave us three directions：

I.Control the saturation level of TS,finding the relationship between the density of holes on nano-silic and protein binding level.

II.Search for reversable parts,such as pH controled parts.

III.We can try to use glycoprotein to build a protein net .

Organization: Beijing Normal University

position:Associate professor

Why do we talk to her?

A number of experiments are needed to test the functions of our silinker.So rich experiences are essential to make sure the experiments are going well.Thus we interviewed Professor Xiaotian Zhang from Beijing Normal University,an expert in molecular biology,especially in protein.We hope to get some professional suggestions on how to do experiments and affirmations of the feasibility and significance of our project.

our takeaways

1.about affirmations of the feasibility and significance：Xiaotian Zhang viewed our project as positive,believing it’s a helpful attempt to make MSN perform a wider range of functions through modification.She gave us a definite reply in respect of similar experimental applications of modification in recent research work.

She also pressed that we should consider more about point-to-point application scenario.When we talked about the potential of silinker,we decided to see it in more detail,Fields of medical treatment,novel nano-material already mentioned,new stakeholders in synthetic medicines and chemical analysis are involved in our project.

2.about how to do experiments：Xiaotian Zhang told us the more comprehensive our experiments are,the more reliable our results are,which also convince people of the feasibility of silinker.Suggestions were given in three aspects.

I.Application conditions：We should first put our experiments in a concrete condition,since different applications also diverse in reaction condition.

II.Literature search：Functions of targeting and affinity can be proved in literature pubished so far.A further discussion is needed on previous methods.We should learn suitable ways to do experiments given in those literatures.

III.Focus and teamwork：It’s unwise to decentralize our team to finish experiments of the four kinds separately.An easy way to incrase efficiency is to gain scale and select useful parts.

Organization: The National Center for Nanoscience and Technology

position：Doctoral supervisor

Why do we talk to him?

Meng Huan is a researcher at the National Center for Nanoscience and Technology, engaged in long-term research in nanopharmacy, drug delivery, nanobiointerfaces, and nanosafety. He has been recognized as a highly cited scientist (Cross-Field) by Clarivate Analytics. He holds multiple patents and has entered the stage of technology transfer.What we want to know:

1.Safety Concerns of Nanosilica as a Drug Delivery Carrier:During our project promotion, we often encounter skepticism about the safety of nanosilica as a drug delivery carrier. There is a prevailing fear of "silicosis" associated with nanosilica, which has led us to urgently investigate the safety of nanosilica as a drug delivery carrier.

2.Advantages and Disadvantages of Nanosilica Compared to Liposomes:We are aware that common drug delivery carriers include liposomes. So, how does nanosilica compare to liposomes in terms of advantages and disadvantages?

Challenges of Protein Modification on Nanosilica and Its Importance:Through extensive literature review, we found that protein modification on nanosilica is a complex and challenging process. We are keen to understand the reasons behind the need for nanosilica protein modification and to verify the difficulties described in the literature.

Simplified Methods for Verifying Protein Targeting Function:In our experimental validation, we encountered some technical difficulties. Therefore, we are seeking simpler methods to verify the protein targeting function.

our takeaways

1.Safety Concerns of Nanosilica as a Drug Delivery Carrier:The skepticism about the safety of nanosilica as a drug delivery carrier is based on generalizations. In reality, the safety of nanosilica depends on its production process, where different techniques can affect the type and density of silanol groups. Nanosilica produced under high-temperature conditions tends to form highly strained three-membered rings, making it extremely unstable and susceptible to ring-opening and free radical generation, posing safety concerns. In contrast, nanosilica produced under low-temperature processes often forms four-membered rings with lower strain, resulting in higher safety levels.

Presently, nanosilica intravenous injection has already received approval from the US FDA, while GRACE, a micrometer-level oral drug, has obtained approval from the European Union. Hence, nanosilica as a drug delivery carrier holds limitless potential for the future.

2.Project Value:

I.Advantages and Disadvantages of Nanosilica Compared to Liposomes:Liposomes are biocompatible carriers with practical applications and concrete instances of use. However, they still face various challenges, as experiments have shown that 20% of drugs loaded in liposomes are released into the environment before reaching the target site, leading to reduced targeting efficiency and significant damage to normal tissues during loading of chemotherapy drugs. Additionally, drug loading capacity and drug solubility in liposomes are correlated. On the other hand, mesoporous silica, as a drug carrier, can improve the solubility of poorly soluble drugs when compared to liposomes. Furthermore, mesoporous silica offers other advantageous properties, such as a large surface area, high porosity, tunable mesoporous structure, and effective enhancement of drug bioavailability.

II.Importance and Advantages of Protein Modification on Nanosilica:Professor Meng Huan explained that unmodified nanosilica colloids have poor stability and tend to aggregate, resulting in structural instability within the in vivo environment. Protein modification serves as an effective strategy to enhance colloidal stability. Therefore, protein modification of nanosilica is necessary to fully utilize its potential.

III.Challenges in Existing Protein Modification:Professor Meng Huan believes that the current protein modification of nanosilica presents difficulties, particularly in terms of high cost and complex processes. Proteins with higher costs require greater investments when exploring modification methods. Our "modular" nanosilica protein modification platform aims to standardize the modification process and simplify complex protein modifications.

IV.Experimental Validation:

(a) pre-experimental validation :He advice us to use fluorescently labeled BSA, cytochrome C, and insulin as models to guide subsequent validation of actual proteins and their real connections.

(b) Fluorescent labeling on nanosilica can be employed to verify correct targeting.

Organization: Beijing Normal University

position:Professor

Why do we talk to him?

In designing how to validate the targeting of our modified mesoporous silica to the cell surface, we have collected a lot of relevant literature and found some feasible methods, but due to the long experimental process and expensive materials, these methods have problems. Therefore, we would like to find relevant experts and ask them how we can effectively verify the targeting of our modified mesoporous silica during the competition.

So we contacted Prof Li Sen, who is a professor at the School of Life Sciences, Beijing Normal University. He has rich working experience and research results in the field of biochemistry and molecular biology, and has led several National Natural Science Foundation projects and Beijing Key Laboratory construction projects, published more than 20 scientific papers, and been granted a number of national invention patents.

our takeaways

1.We can choose cheaper targeted growth factors and antibodies that can target cancer cells, etc. for experimental verification;

2.It is more convenient to culture cancer cells, so it is reasonable to use target cells for validation;

3.Fluorescent labeling can be used for validation by staining the cell itself and the modified mesoporous silica carrier separately, and if the carrier is bound to it, the distribution relationship between the carrier and the cell can be seen, and furthermore, observation after washing can be used to confirm whether the carrier is bound to the surface of the cell or not.

4.In addition to this one can look for more ways to verify the targeting and increase the confidence.

Organization:Beijing Normal University

position:Associate professor

Why do we talk to her?

Shang Yu is an associate professor at Beijing Normal University, specializing in tumor cell biology and anti-cancer drug development. We designed an experiment to verify that our design does not alter the antibody's functionality, ensuring it still effectively targets cells. However, we found that the transferrin, which can target cancer cells according to literature, is too expensive. We hope to consult Professor Shang Yu about the feasibility of our experimental design and identify a more cost-effective yet reliable alternative.

our takeaways

we can compare the cytotoxicity of the drug carrier assembled with slinker linkage to SPG and IgG against tumor cells.