Team:NJTech China/Safety
Safety
Information Sheet
You are invited to participate in the research project "Saving Rose
Plan—Design
and Construction of
Synthetic Yeast-Microalgae Consortia for Biosynthesis of Phenylethanol". The research is being
conducted by team NJTech_China from Nanjing Tech University, China, as part of the iGEM
competition. iGEM is the abbreviation of International Genetically Engineered Machine Competition.
In this competition, students from schools all over the world from up teams and try to solve real
world problems with synthetic biology methods. The purpose of this study is to modify the
Saccharomyces cerevisiae to optimize the phenethylamine pathway of different sources and
expression
modes, improve the activity of phenylacetaldehyde synthase through promoter engineering, and
establish a yeast-microalgae microbial consortia system based on nutritional complementation and
metabolites exchange. The study will help establish a robust, stable, and controllable artificial
microbial consortia system for the green and efficiency phenylethanol biosynthetic.
Your participation involves answering several related
academic questions. The conversation will be recorded. You will be asked of permission of your
identity, words, voice
record, screen record etc.
Your participation in this project is entirely voluntary and
you are free to
withdraw at any time without any consequences. You can also refuse to answer any question. In the
event that you withdraw from the study, your data will be destroyed. To protect your privacy, you
will be anonymized unless you request otherwise. All the information of interviews will be safely
stored on password-controlled computer files and will be destroyed within 6 months after the
competition. Any identifying information will not be shared with anyone else. The data will be used
for "Saving Rose Plan—Design and Construction of Synthetic Yeast-Microalgae Consortia for
Biosynthesis of Phenylethanol" project. A copy of your transcript will be sent to you electrically
upon request.
The finding of our study will be available on
2021igem.org/Team:NJTech_China. If you have
any concerns or questions about this study. you may
contact our team
leader/supervisor at 571688501@qq.com
Interviewer: NJTech_China
Interviewee:
知情书
我们邀请您参加研究项目“拯救玫瑰计划-合成酵母-微藻生物合成苯乙醇混菌体系的设计与构建”。这项研究是由来自中国南京工业大学的NJTech_
China团队进行的，作为iGEM竞赛的一部分。iGEM是国际基因工程机器竞赛的缩写。在这次比赛中，来自世界各地学校的学生组成团队，尝试用合成生物学的方法解决现实世界中的问题。本研究的目的是对酿酒酵母进行修饰，优化不同来源和表达方式的苯乙胺途径，通过启动子工程提高苯乙醛合成酶活性，建立以营养互补和代谢产物交换为基础的酵母-微藻菌群体系。本研究将有助于建立一个稳定、可控的人工菌群体系，实现苯乙醇的绿色高效生物合成。
你的参与包括回答几个相关的学术问题，对话将被录音。您将被要求许可您的身份，文字，声音记录，屏幕记录等。
您参与这个项目是完全自愿的，您可以随时退出而不承担任何后果。你也可以拒绝回答任何问题。如果您退出研究，您的数据将被销毁。为了保护您的隐私，除非您另有要求，否则您将被匿名化。所有的信息将安全存放在有密码控制的电脑文件中，并在比赛结束后6个月内销毁。任何身份信息将不会与任何人共享。该数据将用于“拯救玫瑰计划-合成酵母-微藻生物合成苯乙醇混菌体系的设计与构建”项目。您的受访记录副本将根据您的要求发送电子版到邮箱，我们的研究结果将在2021igem.org/Team:NJTech_China上公布。如果您对这项研究有任何担忧或问题。你可以联系我们的团队
领队/主管: 571688501 @qq.com
采访人:NJTech_China
受访者:
Information Sheet
Q: Describe the goal of your project: what is your engineered organism (or other synthetic biology
product, system, or tool) supposed to do? Please include specific technical details and names of
important parts.
A:
Phenylethanol has been widely used in cosmetics and other fields owing to its rose aroma. Our
project chooses Saccharomyces cerevisiae as the chassis microorganism, optimizes the
phenethylamine
pathway of different sources and different expression modes, overexpresses phenylacetaldehyde
synthase on the strength of promoter engineering, establishes a yeast-microalgae microbial consortia
system based on nutritional complementation and metabolites exchange. The combination with 3D
printing microcapsule embedding technology will eventually guide to establish a robust, stable, and
controllable artificial microbial consortia system for the green and efficiency phenylethanol
biosynthetic.
Q: Which whole organisms, including viruses and cell lines, are you planning to use (or using) in
your project?
A:
(1)Saccharomyces cerevisiae BY4741
BSL-1
(2)E.coli BL21
BSL-1
(3)E.coli DH5alpha
BSL-1
(4)E.coli Rosetta
BSL-1
Q: What risks could these organisms pose to you or your colleagues in the laboratory, or to your
community or the environment if they escape the lab?
A:
Saccharomyces cerevisiae are airborne and able to survive in the air for long periods of
time. In
this way they can contaminate all kinds of other cultures. In addition, allergic people might suffer
from allergio reactions after having been exposed to yeast or its spores. If the transgenic yeast
escapes from the laboratory, the vector and the anti-sequence may be potentially harmful, such as
the uncontrolled spread of the organism or its genetic material.
E.coli DH5alpha is a versatile strain used for general cloning. It is indeed a safe strain.
However,
if the transgenic E.coli escapes from the laboratory, the vectors and the antibiotic
resistance
genes may be potentially harmful, such as the uncontrolled spread of the organism or its genetic
material.
E.coli BL21 and E.coli Rosetta are versatile strains used for protein expression.
It is indeed a
safe strain. However, if the transgenic E.coli escapes from the laboratory, the vectors and
the
antibiotic resistance genes may be potentially harmful, such as the uncontrolled spread of the
organism or its genetic material.
Q: What organisms are you using as chasses in your project?
A:
1. Escherichia coli (give names of all strains you are using): BL21,DH5alpha,Rosetta
2. Yeast (Saccharomyces cerevisiae BY4741)
Q: What risks could your chassis pose to you or your colleagues in the laboratory, or to your
community or the environment if they escape the lab?
A:
Saccharomyces cerevisiae are airborne and able to survive in the air for long periods of
time. In
this way they can contaminate all kinds of other cultures. In addition, allergic people might suffer
from allergio reactions after having been exposed to yeast or its spores. If the transgenic yeast
escapes from the laboratory, the vector and the anti-sequence may be potentially harmful, such as
the uncontrolled spread of the organism or its genetic material.
E.coli DH5alpha is a versatile strain used for general cloning. It is indeed a safe strain.
However,
if the transgenic E.coli escapes from the laboratory, the vectors and the antibiotic
resistance
genes may be potentially harmful, such as the uncontrolled spread of the organism or its genetic
material. E.coli BL21 and E.coli Rosetta are versatile strains used for protein
expression. It is
indeed a safe strain. However, if the transgenic E.coli escapes from the laboratory, the
vectors
and the antibiotic resistance genes may be potentially harmful, such as the uncontrolled spread of
the organism or its genetic material.
Q: As part of your project, are you planning to make / have made new parts or substantively changed
existing parts in the Registry?
A:
Yes.
Q: What experiments will you do with your organisms and parts?
A:
We are interested in the expression of plant-derived phenylacetaldehyde synthase in Saccharomyces
cerevisiae. We obtained the gene sequences of phenylacetaldehyde synthase from petunia, Rose and
Vanda on NCBI website, and had it synthesized by a provider. The synthesized gene was in frame with
the TEF promoter, and it was subsequently overexpressed through the promoter engineering. The
engineered yeasts will be tested in the laboratory. We will also build a mixed bioreactor by 3D
printing, combining yeast and microalgae on bioactive materials in a more stable manner, so as to
build a yeast-microalgae bioreactor model suitable for fermentation and with a good yield.
Q: What risks could arise from these experiments?
A:
Our experiments include relatively basic molecular biology operations such as plasmid extraction,
one-step cloning, PCR, and transformation. The reagents used in these operations may be toxic to the
human body. Although the S. cerevisiae and E.coli are relatively safe chassis cells, they
may be
allergenic. There are ultraviolet lamps for sterilization in the laboratory, and long-term exposure
of personnel under ultraviolet lamps can cause skin diseases and vision damage. Improper operation
of the autoclave may cause steam burns. Sharp tools such as needles and scissors in the laboratory
may stab the operator. High-power electrical appliances such as ovens in the laboratory may have
fire hazards during long-time use.
Q: Are you collecting any data about people, such as their opinions, quotations, health information,
gender, behavior, attitudes, or concerns?
A:
Yes.
Q: Imagine that your project was fully developed into a real product that real people could use. How
would people use it?
A:
In a factory: Using Saccharomyces cerevisiae to produce phenylethanol industrially.
Q: What safety, security or ethical risks would be involved with such a use?
A:
We experimentally demonstrate the concept to explore our GM yeast in factory applications, and we
also consider the risk of accidental release of engineering organisms. Our transgenic yeast may lead
to resistance gene transfer, affecting the growth of wild-type yeast. The media carrying antibiotics
in the experiment are harmful to the environment and may alter the composition of environmental
microorganisms or promote the emergence of super-resistant bacteria. To reduce the environmental and
safety hazards of engineering organisms, we observe all laboratory regulations to prevent accidental
leakage of engineering organisms.
Q: How will experts overseeing your project help to manage any of the risks you identified in this
form?
A:
At Nanjing Tech University, the Laboratory Construction and Management Department
(http://sys.njtech.edu.cn/) is responsible for the
construction and management of the entire
laboratory, laboratory safety, and environmental protection, experimental equipment purchase, and
user management. In our team, Dr. Wenming Zhang is the Principal Investigator whom we consult for
laboratory safety and procedures. He is knowledgeable of the experimental procedures we will be
utilizing, and gives us specific safety advice on these experiments. Guo is also familiar with
laboratory safety procedures in genetic engineering and will ensure the proper use and disposal of
all laboratory materials. Besides, we all have passed the university safety training exam. We
believe our team is qualified to perform these experiments safely.