Safety in the Lab
What biosafety level are our LVV processes?
BSL-2
What comprises the wild type LV genome?
+ssRNA (two copies)
Name the chemical and contact time for liquid LVV biowaste disposal.
0.5% final concentration sodium hypochlorite (10% commercial bleach) for 30 minutes. Then to the biowaste drain.
What is the LVV size range (nm)?
80-100nm
What health care facility do you call if exposed to LVV?
Number Call Medcor (1-800-648-9641) and speak with on-call physician; obtain copy of LVV Medical Protocol front front desk security safe and take to Duke Hospital (3643 North Roxboro St) for evaluation.
What generation are our LVV?
Third-generation self-inactivating LVV
Name the chemical and contact time for LVV surface disinfection.
Soak exposed surface in 6% Steri-Perox for >10 minutes then wipe residue with DECON-Alcohol wipe.
The pH shifts directly pre-transfection from 7.1±0.1 to...
6.7 ± 0.05
True or False: LVV exposure to intact skin does not require clinical treatment since it does not pose a significant health risk.
True, wash exposed skin thoroughly with soap and water for 15 minutes.
What is the vector packaging size?
9kb (8-10kb)
What two additional precautions are required for centrifugation of LVV samples?
Use sealed safety caps on the rotors or buckets and open sealed rotors / buckets in a BSC.
Name the missing plasmid?
(RRE) Rev Response Element
?
(VSVG) Pseudotyping Envelope
(GOI) Gene of Interest
(REV) Regulator of expression of viral proteins
Initiation of post-exposure prophylaxis (antiretroviral drugs) must occur within how many hours of exposure to be beneficial?
72HRS
What virus is rLVV based on (developed from)?
HIV-1
What additional PPE is required for work with LVV (<10L scale)?
Double gloving, leather closed-toed safety shoes or disposable booties, and disposable lab coat. Optional additional PPE includes sleeves (highly recommended) and face shields.
Name all major (7) components within a mature LV particle.
Matrix Proteins (MA)
Reverse Transcriptase
Integrase
Protease
+ssRNA (two copies) viral or recombinant genome
Nucleocapsid (NC)
Capsid (CA) (p24)
How many recombination events are required in order to produce RCL?
Four recombination events between plasmids without homology and pick-up of a promoter to complement SIN deletion (deletions in the 3' LTR of the viral genome disrupted the promoter/enhancer activity of the long terminal repeat) would be required to produce RCL.
What does VSV-G stand for?
Vesicular Stomatitis Virus Glycoprotein
What additional PPE is required for work with LVV (>10L scale)?
Disposable lab suit (coveralls / bunny suits) and disposable shoe covers.
Name all 9 proteins encoded within the wtHIV-1 genome.
Gag
Pol
Env (replaced by pseudotyping)
Tat (deleted to form Gen3)
Rev
Nef (deleted to form Gen2)
Vif (deleted to form Gen2)
Vpu (deleted to form Gen1)
Vpr (deleted to form Gen2)