Team:NJU China/Acknowledgement

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<h1>Attribution<span>Here we attribute work done by others and by ourselves. We distinguish work done by the team from work done by others, including the 3M lab group, advisers, instructors, graduate students, and postgraduate masters students.</span></h1>
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<h1>iGEM 2013 Basic Safety Form <span>Team name: NJU-China</span></h1>
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</br>    <li><a href="https://2013.igem.org/Team:NJU_China/Judging criteria">Judging criteria</a>
 
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    <li><a href="https://2013.igem.org/Team:NJU_China/Acknowledgement">Acknowledgement</a>
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</br>    <li><a href="https://2013.igem.org/Team:NJU_China/Safety">Safety form</a>
    
    
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<p>Work done by team</BR></BR>
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<p>
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　　Contributed innovative ideas for the project(All team member)</BR>
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Virus-related safety</br>
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　　Erformed all molecular experiments ( plasmid construction, Luciferase, RT-PCR)(WENG Mingxi, NIU Yuchen, ZHOU Yu, WEI Xiuqing, DAI Yimei, CHEN Xi, XIONG Aoli, WANG Wei, GONG Fei, CAI Yusheng)</BR>
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Our project might raise some safety concern for we used part of the Hepatitis B virus and Rabies virus coat protein in our targeting fusion protein. However, what we are using are only short peptides from these two viruses, which lack the central part for viral replication. Apart from that, these two peptides are widely used in lab and they won’t pose any danger to researchers [1].</br>
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　　Performed all culture experiments (transfection, growth, exosome collection) (mainly done by WANG Wei, WEI 　　Xiuqing, DAI Yimei, CHEN Xi, XIONG Aoli, SUN Yiyang, NIU Yuchen, WENG Mingxi, GONG Fei)</BR>
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Details of the peptides we are using are stated below:</br>
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　　Performed all biochemical experiments (CHEN Xi, DAI Yimei, WEI Xiuqing, NIU Yuchen)</BR>
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</br>
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　　Preformed all mice experiments (SUN Yiyang, WEI Xiuqing, WANG Wei, DAI Yimei, CAI Yusheng)</BR>
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Pre-S1 from the Hepatitis B virus (HBV)</br>
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　　Modeling (GONG Fei, WANG Siqi)</BR>
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HBV is double-stranded DNA virus and it is among the Risk group 3. We are in biosafety level 2 lab so we are not legally allowed to deal with the virus directly. And we never deal with the real virus. What we are using is only the gene of pre-S1(GenBank: AJ131540.1).</br>
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　　Designed Wiki (ANN Hongrui, GONG Fei)</BR>
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HBV coat protein is composed of three proteins, small(S), middle(M) and large(L). Pre-S1 is a peptide at the N-terminus of the large coat protein, which can mediate specific recognition of the hepatic cells[2]. We obtain the sequence from GenBank: AJ131540.1 and synthesized the sequence by Invitrogen. Then we cloned the sequence into the pcDNA 3.1(+) vector. We confirmed that no other viral gene is contained in our plasmid by sequencing. Since we use only the pre-S1 from the HBV genome, it won’t pose any danger to lab members or public.
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　　Planed and Organized human practice ( All team member)
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RVG from the Rabies virus</br>
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Rabies is single-stranded RNA virus. What we are using is a short piece of peptide from the glycoprotein of Rabies virus.</br>
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We get the sequence from the nature paper[3] and synthesized it from Invitrogen. Then we cloned the sequence into pcDNA 3.1(+) vector. We confirmed that no other viral gene is contained in our plasmid by sequencing. Since we use only the RVG from the Rabies virus genome and it lacks the replication ability, it won’t pose any danger to lab members or public.</br>
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</br>
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Reference:</br>
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[1] Ma, Y., R.J.M. Nolte, and J.J.L.M. Cornelissen, Virus-based nanocarriers for drug delivery. Advanced Drug Delivery Reviews, 2012. 64(9): p. 811-825.</br>
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[2] Yan H, Zhong G, Xu G, et al. Sodium taurocholate cotransporting polypeptide is a functional receptor for human hepatitis B and D virus[J]. Elife, 2012, 1.</br>
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[3] Alvarez-Erviti L, Seow Y, Yin H F, et al. Delivery of siRNA to the mouse brain by systemic injection of targeted exosomes[J]. Nature biotechnology, 2011, 29(4): 341-345.</br>
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<p>Support of advisers</BR></BR>
 
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　　Basic training of students for techniques of molecular and cell biology (WANG Xueliang, LIANG Hongwei, LIU Yanqing)</BR>
 
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　　Technical support on mice experiments(ZHANG Yujin)</BR>
 
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　　Technical support with confocal microscopy (WANG Nan)</BR>
 
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　　Discussing the results and editing the wiki (all advisers)</BR>
 
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　　Soliciting funds for the project (all advisers)</BR>
 
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<p>Support received from others</BR></BR>
 
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　　We have gained much information about iGEM from other teams: WU Xin from XMU, ZHANG Sitao from USTC, XUE Angli from ZJU.</BR>
 
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　　Also, CHEN Shuobing from Peking University has given us precious advice about our project. Professors from NIBS have given us much information about the pre-S1.  </BR>
 
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　　Moreover, our adorable classmates, HUA Yilei and WEI Yu have given us much help in Logo design and presentation, respectively.</BR>
 
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<p>Materials received from 3M lab</br></BR>
 
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　　HEK 293T Cell line</br>
 
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　　HepG2 Cell line</br>
 
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　　Plasmids used as template</br>
 
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<a href="https://static.igem.org/mediawiki/2013/4/4f/NJU-IGEM_Biosafety_Form_Part_2_HEK293.pdf">IGEM_Biosafety_Form_Part_2 HEK293</a>
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<p>Material received from others</BR></BR>
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</br>
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　　HBsAg Mice Model from CUST</BR>
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<a href="https://static.igem.org/mediawiki/2013/c/ca/NJU-IGEM_Biosafety_Form_Part_2_HepG2.pdf">IGEM_Biosafety_Form_Part_2_HepG2</a>
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</br>
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<a href="https://static.igem.org/mediawiki/2013/1/13/NJU-IGEM_Biosafety_Form_Part_2_Lamp_2b-RVG.pdf">NJU-IGEM_Biosafety_Form_Part_2_Lamp_2b-RVG</a>
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<p>Mathematical model</BR></BR>
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</br>
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　　Inspired by iGEM Team Slovenia 2012 </BR>
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<a href="https://static.igem.org/mediawiki/2013/d/db/NJU-IGEM_Biosafety_Form_Part_2_Lamp_2b-preS1.pdf">NJU-IGEM_Biosafety_Form_Part_2_Lamp_2b-preS1</a>
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Revision as of 00:23, 28 September 2013

<!DOCTYPE html> NJU_China

iGEM 2013 Basic Safety Form Team name: NJU-China

Virus-related safety
Our project might raise some safety concern for we used part of the Hepatitis B virus and Rabies virus coat protein in our targeting fusion protein. However, what we are using are only short peptides from these two viruses, which lack the central part for viral replication. Apart from that, these two peptides are widely used in lab and they won’t pose any danger to researchers [1].
Details of the peptides we are using are stated below:

Pre-S1 from the Hepatitis B virus (HBV)
HBV is double-stranded DNA virus and it is among the Risk group 3. We are in biosafety level 2 lab so we are not legally allowed to deal with the virus directly. And we never deal with the real virus. What we are using is only the gene of pre-S1(GenBank: AJ131540.1).
HBV coat protein is composed of three proteins, small(S), middle(M) and large(L). Pre-S1 is a peptide at the N-terminus of the large coat protein, which can mediate specific recognition of the hepatic cells[2]. We obtain the sequence from GenBank: AJ131540.1 and synthesized the sequence by Invitrogen. Then we cloned the sequence into the pcDNA 3.1(+) vector. We confirmed that no other viral gene is contained in our plasmid by sequencing. Since we use only the pre-S1 from the HBV genome, it won’t pose any danger to lab members or public.

RVG from the Rabies virus
Rabies is single-stranded RNA virus. What we are using is a short piece of peptide from the glycoprotein of Rabies virus.
We get the sequence from the nature paper[3] and synthesized it from Invitrogen. Then we cloned the sequence into pcDNA 3.1(+) vector. We confirmed that no other viral gene is contained in our plasmid by sequencing. Since we use only the RVG from the Rabies virus genome and it lacks the replication ability, it won’t pose any danger to lab members or public.

Reference:
[1] Ma, Y., R.J.M. Nolte, and J.J.L.M. Cornelissen, Virus-based nanocarriers for drug delivery. Advanced Drug Delivery Reviews, 2012. 64(9): p. 811-825.
[2] Yan H, Zhong G, Xu G, et al. Sodium taurocholate cotransporting polypeptide is a functional receptor for human hepatitis B and D virus[J]. Elife, 2012, 1.
[3] Alvarez-Erviti L, Seow Y, Yin H F, et al. Delivery of siRNA to the mouse brain by systemic injection of targeted exosomes[J]. Nature biotechnology, 2011, 29(4): 341-345.

IGEM_Biosafety_Form_Part_2 HEK293
IGEM_Biosafety_Form_Part_2_HepG2
NJU-IGEM_Biosafety_Form_Part_2_Lamp_2b-RVG
NJU-IGEM_Biosafety_Form_Part_2_Lamp_2b-preS1

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