Caco-2 Transporter Knockout Cell-Based Assays

Service Cell Line Transporter Technology Membrane Type Probe Substrate Read Out Reference Inhibitor Positive Control Dynamic Range
Caco-2 BCRP knockout substrate assessment BCRP
Monolayer Assay
estrone-3-sulfate
Caco2 MDR1 knockout substrate assessment MDR1/P-gp
Monolayer Assay
digoxin
Caco-2 MRP2 knockout substrate assessment MRP2
Monolayer Assay
CDCF

Description

Substrate assessment with efflux transporter knockout cells

SOLVO expanded its monolayer substrate assay portfolio with transporter specific Caco2 (C2BBe1 subclone) knockout cells. Transporter knockout Caco2 cells are a good method for determining the potential impact of specific efflux transporters in the intestine on the absorption and disposition of a drug.

Transporter knockout cell lines can be used to identify specific drug-transporter interactions by comparison of drug transport between the wild-type (WT) and the knockout cell lines, without dependence on chemical inhibitors. Transporter specific knockout strains of the Caco2 cell line were developed by Sigma-Aldrich (Sigma, St Louis MO). Functional knock out Caco2 cells were developed by utilizing CompoZr® Zinc Finger Nucleases (ZFNs) [1,2]. The resultant MDR1, BCRP and MRP2 knockout cells demonstrated disruption of gene sequence as well as complete loss of transporter function in bidirectional transport assays out to at least 40 passages post-ZFN genomic modification.

These cell lines have been characterized at SOLVO and proven to be an alternative tool for elucidating drug-transporter interactions. Model compounds, known to be substrates for these efflux transporters such as: digoxin, estrone 3-sulfate, and CDCF have been used, and in each case, the efflux ratio of the compound decreases close to unity in the appropriate knockout cell line versus the WT cell line. Characterization data is available upon request.

Substrate assessment assay setup

References:

1. Carroll D (2011) Genetics 188; 773-782
2. Pratt, J et al. , (2012) Current Protocols in Toxicology 52. 23.2.1-23.2.22