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Application

Knock-out cell lines and their use in signaling pathways

Abcam

Jan 28, 2022

Abstract

Knock-out (KO) cell lines are powerful tools to study disease mechanisms, such as the effect of gene disruption on downstream pathways. In this application note, we highlight the effectiveness of Abcam KO cell lines that can be used to investigate signaling pathways, in this case, the TRAIL pathway. TRAIL (TNF-related apoptosis-inducing ligand) is a protein cytokine that causes apoptosis through binding to several death receptors. These receptors have gained substantial interest as cancer cells are significantly more sensitive to TRAIL-induced apoptosis than normal cells. We used a KO cell line of a TRAIL death receptor, DR5, in HeLa cells (ab264922) to demonstrate that KO attenuates the downstream apoptotic pathway initiated by TRAIL. Upon activation of the pathway, the signal is propagated through a series of effector caspases, including caspase 3. Therefore, we included a small-molecule caspase 3 inhibitor, Q-DEVD-OPh, as a positive control. Demonstrating the advantage of a complete genetic KO, the DR5 KO line had a greater effect on inhibiting the downstream signaling pathway than Q-DEVD-OPh at the concentrations used.


Application File

KO-cell-lines-TRAIL-appnote.pdf

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