The “ABCS” (Adipocyte for Breast Cancer Surveillance) project aims to detect breast cancer through engineered adipocytes. Our RADAR system is designed to express Humanized Gaussia Luciferase (hGluc) when detecting the upregulated expression of target genes (LIF, PLOD2) in response to the stimulation of tumor-associated cytokines (CXCLs, PAI-1). With the addition of the substrate of Gaussia Luciferase, coelenterazine, RADAR system allows the detection of CXCLs and PAI-1 directly by observing the presence of luminescence. By packaging this RADAR system in rAAV and infecting into adipocytes, such engineered adipocytes could be used to detect breast cancer cells that secrete CXCLs and PAI-1 in vivo.
The sensor sequence is the core of “RADAR” system. (Please see Description for detail information.) Sensor 1 and Sensor 2 are reported in previous published literature and used for our proof of concept experiments. Sensor_hLIF and Sensor_hPLOD2 are designed by us and used to detect the expression of target genes: LIF and PLOD2.
When sensor binds to its target RNA, it recruits the ADAR enzyme, which modifies the AUG stop codon in the sensor sequence to initiate downstream gene translation. mCherry is used to demonstrate delivery efficiency, and EGFP or Gluc is the output to validate the ability of sensor to detect the upregulated expression of target genes. The “h” indicate human (Homo sapiens).
We have designed and registered 12 basic parts and 8 composite parts.
List of New Basic Parts
Composite Parts
No.
Title
Diagram
Type
Length (bp)
List of New Composite Parts
No.
Title
Diagram
Type
Length (bp)
