eBook: Cell and Gene Therapy - 20

Figure 2. Dengue virus type 2 strain New Guinea C titer comparison. Vero parental cells and Vero.STAT1 KO cells were seeded and infected with
Dengue II New Guinea C virus. Viral supernatants were harvested at day 7 post infection and then titered by further infecting WT Vero cells in
the indicated serial dilutions. (A) Dengue II viruses within the cells were analyzed by immunofluorescence staining and high content microscopy
imaging. (B) The 50% tissue culture infective dose (TCID50
) were also calculated (n=9). (C) Additionally, RT-PCR shows 30-fold increase in Dengue
virus type 2 genomes produced in STAT1 KO Vero cells compared to the parental Vero cells.
used to infect MDCK.STAT1 KO and MDCK cells, and
high-content imaging and RT-PCR was performed
as before (Figures 3A-C). Similar to the gene-edited
Vero cells, a 10-fold increase in Influenza A virus-
positive cells and a 2-fold increase in viral genomes
was observed in KO cell line as compared to the parental
MDCK cells.
Finally, to examine the 293.STAT1 BAX KO cell line's
virus-producing competence, we infected both
the knockout and wild type HEK 293 cell lines with
GFP-expressing Sendai virus (ATCC® VR-105™) and
harvested supernatants 48 hours post-transduction.
As before, serial dilutions of the viral supernatants
were used to infect the knockout and wild
type cell lines, and high-content imaging and RTPCR
was performed (Figures 4A-C). In line with the
STAT1 KO cell lines, we saw a more than 5-fold increase
in Sendai virus-positive cells in the KO cell
line as compared to the parental HEK 293 cells. To
examine the 293.STAT1 BAX KO cell line's capacity
of producing AAV, we transfected the cells with an
AAV5 viral vector (ATCC® VR-1523™) and collected
20

https://www.atcc.org/products/vr-105 https://www.atcc.org/products/vr-1523

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