Team:Mingdao/Safety

Mingdao iGEM Lab Safety Regulations

All personnel receives a tour and are informed of the rules upon entering the lab, the main rules include:

1. No running, sudden moves within the lab.

2. No eating within the premises of the lab.

3. All experiment waste must be thrown in a special bin then put through a machine for sterilization.

4. All pipettes must be sanitized after use.

5. All equipment use must follow the using manual.

6. The mosquito cage must always be double sealed.

7. A bottle of ethanol must always be around when handling the mosquitoes.

8. To enter the P2 lab, all personnel must be thoroughly sanitized, must wear full lab suits, remove shoes and watches, and pass through two doors.

Q&A Regarding Project Safety

Q1:

Which whole organisms, including viruses, are you planning to use or using in your project?

A1:

1. C6/36 cell line from Aedes albopictus which is bought from Bioresource Collection and Research Center in Taiwan.

2. Drosophila S2 cell line is given by Prof. Kuang-Hui Lu at National Chung Hsing University.

3. E. coli DH5alpha competent cells are purchased from the biotech company of Geneaid Inc.

4. Aedes aegypti adult mosquitoes are bred by the mosquito lab of Prof. Wu-Chun Tu at National Chung Hsing University. The mosqutioes are used to test the blood sucking behavior under strict regulation and instruction by the supervisor of Dr. Hou-You Tzeng from Prof. Tu's Lab. The experiment are performed by human hands wearing air-permeable gloves to attract the mosquitoes and observing mosquitoes' behavior.

Q2:

What risks could these organisms pose to you or your colleagues, or to your community or the environment if they escape the lab?

A2:

1. The mosquito could fly around drawing blood from people. The mosquito experiments are performed in a quarantined mosquito room with an extremely high level of safety regulation with experienced mosquito handlers from Prof. Wu-Chun Tu's lab at National Chung Hsing University, thus, the odds of a mosquito escaping is extremely low.

2. The cell lines and E. coli strain are recognized as Biosafety Level 1 by U.S. Public Health Service Guidelines and pose less risk to humans.

Q3:

What risks could your chassis pose to you or your colleagues, or to your community or the environment if they escape the lab?

A3:

E.coli DH5 alpha, the mosquito C6/36 cell line and Drosophila S2 cells will not pose a threat to anyone since they are labelled as biosafety level one. Such cells can't cause consistently diseases in immunocompetent adult humans and present minimal hazard to the environment.

Q4:

What experiments will you do with your organisms and parts?

A4:

We would use C6/36 cell line from Aedes albopictus to check the pathways in a mosquito cell.( The cells are bought from Bioresource Collection and Research Center in Taiwan)

We will also be testing the Aedes aegypti's circadian rhythms to know when they will draw blood. This experiment would be done by holding a limb outside the cage and determine how long they would be drawn over, so there would be no real blood in this experiment(The Aedes aegypti mosquitoes are bred by a the mosquito lab of Professor Wu-Chun Tu under the regulation of Center for Environmental Protection and Occupational Safety and Health,NCHU.)

Q5:

What risks could arise from these experiments?

A5:

The main hazard this experiment could create is only if the mosquitoes escape, but that would be quite impossible since the cage is impenetrable for the mosquito.

Q6:

What safety, security or ethical risks would be involved with such a use?

A6:

It is possible that people wouldn't like being bitten by a mosquito. And though extremely unlikely, the mosquito might escape from its container and go around biting people.

Q7:

How will experts overseeing your project help to manage any of the risks?

A7:

We have worked under the supervision of National Chung Hsing University's mosquito lab, where they have safety regulations specifically in mosquito biosafety. They also have a legal permit on the mosquito experiments.

Q8:

What rules or guidance cover your work which could help to manage any of the risks you identified in the last section?

A8:

The main regulations regarding bio-safety and bio-security is published by CDC Taiwan, the main precautions are sanitizing, surveillance, reporting, biosafety inspections of high-containment laboratories, biosafety mechanism registration, laboratory biosafety education and training, and laboratory bio-risk management.

Safety Guide for the Lab of Genetic Engineering, Taiwan National Science Commitee (words in Chinese)

Laboratory Biosafety Regulations in Mingdao High School (words in Chinese)

Q9:

How does your bio-safety training help you identify the risks in the project.

A9:

The bio-safety training all members received helps us to be extra cautious when handling the mosquitoes, since it could fly away and spread diseases found in our area. For safety measures, we always keep the seal blocked twice over and keep a bottle of ethanol around the cage (to kill any mosquito that might happen to escape). We have been asked multiple times how we are going to stop the mosquitoes from escaping and spreading diseases. We came up with two possible solutions, the first is a simple yet effective one that requires only plucking off the wings, thus stopping the mosquitoes from spreading. The second solution is quite difficult since it requires bio-engineering the mosquitoes to stop them from giving birth, so we find this one not that convincing.

Part Information

Part: BBa_K2543000

Name: Ac5 promoter / pSB1C3

Natural function of part: Actin promoter

Source: PCR from pAc5.1/V5-His Vectors

Use: To constitutively express GFP

Species: Drosophila melanogaster

Risk Group: No (Animals and plants have no Risk Group)

Disease: No

Part: BBa_K2543001

Name: GAM1 promoter / pSB1C3

Natural function of part: Antimicrobial peptide promoter

Source: PCR from gDNA of Aedes aegypti mosquitoes which are bred in a mosquito lab of National Chung Hsing University

Use: To regulate AMP expression

Species: Aedes aegypti

Risk Group: No (Animals and plants have no Risk Group)

Disease: No

Part: BBa_K2543002

Name: SV40 poly A / pSB1C3

Natural function of part: RNA processing

Source: PCR from pAc5.1/V5-His Vectors

Use: To help gene expression

Species: Simian virus 40

Risk Group: Bio Safety Level 2 according to U.S. Public Health Service Guidelines

Disease: a potent DNA tumor virus

Part: BBa_K2543006

Name: Human CD4 / pSB1C3

Natural function of part: T cell biomarker and induce T cell activation signaling

Source: IDT gene synthesis

Use: To response to gp120 protein of HIV

Species: Homo sapiens

Risk Group: No (Animals and plants have no Risk Group)

Disease: No

Part: BBa_K2543007

Name: Drosophila Toll / pSB1C3

Natural function of part: Toll signaling to induce antimicrobial peptide (AMP)

Source: IDT gene synthesis

Use: To induce AMP promoter activity

Species: Drosophila melanogaster

Risk Group: No (Animals and plants have no Risk Group)

Disease: No

Photos of the LAB

Eating and Drinking are not Allowed in the Lab.

The Laminar Flow Clean Bench in the BIOLAB, MDHS.

Warning Signs are Posted in the LAB.

The Emergency Eyewash in the BIOLAB, MDHS.

Autoclave

The BIOLAB of Mingdao High School.

Performing Experiments with a Pipette while Wearing a Lab Coat and Gloves.