Project Safety Issues
Our project raises several safety concerns, primarily for the researcher and to a small degree, the public. Several of our project components have been designated as Biosafety Level 2 (BSL2).
First, our project uses the tat cell-penetrating peptide, originally found in HIV-1. Tat has the ability to cross cell membranes, and can be used for cellular uptake of assorted molecular cargo, if the cargo is bonded to the peptide. In the past, tat has been used to deliver such products as GFP and DNA to cells. Because of the "infectious" characteristics of the tat peptide, it has been designated at a BSL2. While working with the peptide, the researcher must be careful to cover exposed skin areas (wear gloves and long sleeves, or a lab coat), and wear goggles. Work with the peptide must be in a proper microbiological hood. For more information on tat safety guidelines, please see this page from the University of Chicago.
Second, the glioblastoma cells we are using come from human cancer tissue. Because the cells are human-derived, they are automatically considered BSL2.
Third, the BV-2 microglial cells we are using are derived from mice. However, these microglia have been immortalized by genetic modification. As a result, they are considered BSL2.
The mammalian culture cell lab in which we work is a BSL2 facility with equipment and protocols that follow BSL2 guidelines, minimizing safety concerns for the researcher.
Local Biosafety Review Board
At Purdue University, the organization in charge of biosafety is the Radiological and Environmental Management (REM) Department. They work in cooperation with the Institutional Biosafety Committee (IBC). For work with recombinant DNA, a protocol must be reviewed and accepted by them to ensure safety.
We are still in the process of getting our protocol approved. However, we feel confident it will be approved because we have the proper safety training, equipment, and protocols for this safety level.
The part we submitted this year was part BBa_K217000, which codes for the constitutive production of the tat-GFP fusion protein. We have documented on the part's main page the safety considerations concerning the part. Here is a link to the registry page for the part: our part's main page