Team:NJU China/Virus-related safety

From 2013.igem.org

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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