An article published this year in “ONCOTARGET” using our Annexin
V-Dy634 apoptosis detection kit, by our customers from Unidad de
Diagnóstico Molecular y Celular del Cáncer, Centro de Investigación del
Cáncer-IBMCC (USAL-CSIC), and IBSAL, Instituto de Investigación Biomédica de
Salamanca, Spain, in the study of how the CRISPR-Cas9 system efficiently
reverts the tumorigenic ability of BCR/ABL in vitro and in a xenograft model of
chronic myeloid leukemia. Congrats and Thanks.
Summary:
CRISPR/Cas9 technology was used
to abrogate p210 oncoprotein expression in the Boff-p210 cell line, a pro-B
line derived from interlukin-3-dependent Baf/3, that shows IL-3-independence
arising from the constitutive expression of BCR-ABL p210. Using this approach,
pools of Boff-p210-edited cells and single edited cell-derived clones were
obtained and functionally studied in vitro. The loss of p210 expression in
Boff-p210 cells resulted in the loss of ability to grow in the absence of IL-3,
as the Baf/3 parental line, showing significantly increased apoptosis levels.
Notably, in a single edited cell-derived clone carrying a frame-shift mutation
that prevents p210 oncoprotein expression, the effects were even more drastic,
resulting in cell death. These edited cells were injected subcutaneously in
immunosuppressed mice and tumor growth was followed for three weeks.
BCR/ABL-edited cells developed smaller tumors than those originating from
unedited Boff-p210 parental cells. Interestingly, the single edited cell-derived
clone was unable to develop tumors, similar to what is observed with the
parental Baf/3 cell line.
CRISPR/Cas9 genomic editing
technology allows the ablation of the BCR/ABL fusion gene, causing an absence
of oncoprotein expression, and blocking its tumorigenic effects in vitro and in
the in vivo xenograft model of CML. The future application of this approach in
in vivo models of CML will allow us to more accurately assess the value of
CRISPR/Cas9 technology as a new therapeutic tool that overcomes resistance to
the usual treatments for CML patients.
Reference:
Product link:
DY634 Annexin V Apoptosis Detection Kit with PI: http://www.immunostep.com/apoptosis-tools/3736-anxvkdy-100t.html
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