59
185
Lentiviruses or Retroviruses productions
136
Transductions in eucaryotic cells
32
Users
Examples of publications including
results partly obtained thanks to the
GIGA-Viral Vectors platform
Condé C, Rambout X, Lebrun M, Lecat A, Di
Valentin E, Dequiedt F, Piette J, Gloire G, Le-
grand S. The inositol phosphatase SHIP-1 inhi-
bits NOD2-induced NF-kappaB activation by
disturbing the interaction of XIAP with RIP2.
PLoS One 2012;7:e41005.
Lecat A, Di Valentin E, Somja J, Jourdan S,
Fillet M, Kufer TA, Habraken Y, Sadzot C,
Louis E, Delvenne P, Piette J, Legrand-Poels
S. The c-Jun N-terminal Kinase (JNK)-binding
Protein (JNKBP1) Acts as a Negative Regu-
lator of NOD2 Protein Signaling by Inhibi-
ting Its Oligomerization Process. J Biol Chem
2012;287:29213-26.
Rubio N, Coupienne I, Di Valentin E, Heirman
I, Grooten J, Piette J, Agostinis P. Spatiotem-
poral autophagic degradation of oxidatively
damaged organelles after photodynamic
stress is amplifed by mitochondrial reactive
oxygen species. Autophagy 2012;8.
Sabatel H, Di Valentin E, Gloire G, Dequiedt
F, Piette J, Habraken Y. Phosphorylation of
p65(RelA) on Ser(547) by ATM Represses
NF-kappaB-Dependent Transcription of Spe-
cifc Genes after Genotoxic Stress. PLoS One
2012;7:e38246.
Viral Vectors
Created in 2012, the Viral Vector platform produces customized lentiviral or retroviral vectors whose
production requires to work in Biosafety 2 or 3 labs (BSL2 or BSL3). This plateform also assume ULg
scientists training in order to have an access to the BSL2 or BSL3 (A3) laboratories of the GIGA. This
allow scientists to work into a safe environment with diferent virus or viral vectors within the respect
of biosafety rules.
Depending on the request, the platform’s staf can help for the design and the cloning into a retro/len-
tiviral plasmids. These plasmids then are used for production of standard vectors (universal or specifc
promoters, multicistronic constructs , tagged vectors), imaging Vectors (eGFP, Luciferase, RFP, mCherry,
BFP), RNA interference vectors for stable knock-down (shRNA RNAi-vectors polIII promoters or indu-
cible promoters) and alternative pseudotyping (VSV-G, GalV, Measles, Ampho and eco, 10A1 MLV). The
viral vectors are produced within two weeks. A titration is done by RTqPCR and the staf assure a titer
of a minimum 10^6 transduction units per mililiters (TU/mL) or higher. If needed the produced vectors
can be be purifed and concentrated in order to reach up to10^8 TU/mL. Cotransfection with minimum
3 diferents plasmids is used to produce non replication competent retroviral particles (non-RCR), this
techniques narrows the chance of recombinaison between the plasmids and prevent the production of
hypothetical wild type viruses.
The platform can also transduce cells with these viral vectors. After selection and an amplifcation of the
transduced cells, supernatants are checked for the absence of RCR for allowing to use cells into BSL1.
The entire process takes approximately 4 weeks. All the produced vectors and cells can be used in vitro
and in vivo (in animals).
Curently the platform is working on the developpment of recombinants Adeno associated virus (rAAV)
vectors productions. AAV are not currently known to cause human disease and consequently the virus
causes a very mild immune response. rAAV can transduce both dividing and non-dividing cells. Those
rAAV vectors are specifcaly design to allow an overexpression of the gene of interest, generate a knock
or a point mutation in on both alleles through an homologous recombinaison. This technic will be mainly
used to introduce a stop codon or a specifc mutation on the desire location.
Contact
Emmanuel Di Valentin
viralvectors.giga@ulg.ac.be
www.giga.ulg.ac.be/viralvectors