Home › Cell Biology › Utilizing Functional Genomics Screening to Identify Potentially Novel Drug Targets in Cancer Cell Spheroid Cultures
Cell Biology JoVE (Open Access) Citable · DOI

Utilizing Functional Genomics Screening to Identify Potentially Novel Drug Targets in Cancer Cell Spheroid Cultures

DOI: 10.3791/54738-v
What you'll learn
  • ✓ Perform reverse transfection of cancer cells
  • ✓ Culture BT474 cells in spheroid format
  • ✓ Acquire automated images of spheroids
  • ✓ Analyze functional genomic screening results
Protocol

Biopharma Insights Identifying novel drug targets that transition from pre-clinical testing to human trials is a scientific priority. To that end, here we describe a functional genomics approach for examining the impact of gene depletion on cancer cell line spheroids, which more appropriately model human cancers in vivo.

Difficulty
intermediate
Total time
~1 week per screen
Model organism
BT474 cancer cell line
Biosafety
BSL-1

Steps

1
Reverse transfect BT474 cells

Perform reverse transfection of BT474 cells to introduce gene-specific siRNAs or shRNAs.

▶ 01:17
2
Acquire images automatically

Use automated microscopy to capture images of the spheroids at the designated time points.

▶ 03:51
3
Analyze oncogenic dependencies

Examine the imaging data to identify genes whose depletion alters spheroid growth or viability.

▶ 05:23
💬 Comments coming soon