In accordance to the iGEM safety rules, our team used a cellular
organism classified in the Risk Group 1. For the purpose of cloning and
expression, Escherichia coli K-12-strain derivates were primarily
applied in the lab as the main experimental subjects. Standard plasmids
with standard antibiotic resistance markers were designed to enhance
functions of recombinants selection. To ensure the safety of ourselves and
the project, we worked in a BSL-1 (Biosafety type 1) laboratory,
following standard safety protocols (e.g. masks, gloves, googles, and
protective clothing). The lab also includes the “clean-to-dirty” workflow
area, which means that lab staffs do not return to a “clean” area after
working in a “dirty” area. For instance, entering PCR room is forbidden
after working with DNA isolation or bacterial cultures in DNA room.
One of our project’s part which raised concern about biosecurity is the
synthetic hand-made Affitoxin (i.e. the binding domain of AB diphtheria
exotoxin) design. The details regarding this part would be discussed in
this page ( http://parts.igem.org/Part:BBa_K2607000). The fact
that this part originates from the modification of diphtheria toxin raises
concern about safety and security, including the possibilities of dual-use
research. Regarding safety issues, the main purpose of the creation of this
part is to eliminate the toxic domain of the exotoxin, leaving only the binding
domain to be used to test our chimeric HB-EGF/Tar receptor. However, this
condition still leaves a security issue regarding dual-use possibilities.
Additionally, the creation of this part, if successful, means that we have
only isolated the necessary domains needed for binding assays. The full
sequence could be used to construct another chimeric protein in which the
Affitoxin is linked to a more harmful toxin or material and used to
deliver that harmful part into the cytoplasm of human cells containing
HB-EGF surface receptor. To handle this, our team are required to comply
with rules of iGEM safety committee to be allowed to not publish the full
sequence in the iGEM UI 2018 Wiki pages.
