eBook: Cell and Gene Therapy - 22

Figure 4. Sendai virus (SeV) and AAV5 titer comparison. (A) TCID50 of GFP SeV supernatants produced by WT parental and 293.STAT1 BAX KO
cells. Cells were infected with GFP SeV at an MOI of 0.01. Supernatants were collected 48 hours after infection and used to reinfect WT HEK 293
cells at the indicated dilution. (B) Cells were imaged 48 hours post GFP SeV infection, Scale bar=1000 μm. (C) TCID50
of GFP SeV supernatants
produced at 48h post infection were calculated (n=3). (D) Cells were transfected with AAV5 viral vector. The supernatants were collected 48
hours after transfection and used to re-infect WT HEK 293 cells. Droplet digital PCR quantification of AAV5 viral genomes indicated a 1.8-fold
increase in viral genomes produced in 293.STAT1 BAX KO cells compared to the parental cells at 72h post-infection.
pression of STAT1 and BAX. All 3 cell lines displayed
increased viral production capacity. These highly
validated cell lines were thoroughly characterized
for viral production phenotype, stability of the
STAT1 and STAT1/BAX double knockout genotype,
and absence of off-target mutations.
ATCC's cell lines for enhanced viral production have
broad applicability for bioproduction in basic research
and the pharmaceutical industry. Because
of their ability to yield higher virus titers than their
wild-type counterparts, these enhanced virus-producing
cell lines have the potential to significantly
reduce the costs associated with generating viral
vaccines and gene therapies.
References
1. Horvath CM, Darnell JE Jr. The antiviral state induced
by alpha interferon and gamma interferon requires
transcriptionally active Stat1 protein. J Virol 70: 647650,
1996. PubMed: 8523587
2. Meraz MA, et al. Targeted disruption of the Stat1 gene
in mice reveals unexpected physiologic specificity
in the JAK-STAT signaling pathway. Cell 84: 431-442,
1996. PubMed: 8608597
3. Cuconati A, et al. Bak and Bax function to limit
adenovirus replication through apoptosis induction. J
Virol 76(9): 4547-4558, 2002. PubMed: 11932420
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