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Reveal Hidden Details of CRISPR P53 Knockout Cells Using an Adapted Cell Painting Assay

Discover how advanced near-infrared imaging reveals clearer cellular insights

Tumor protein p53 plays a pivotal role in regulating cell health—yet mutations in the p53 gene are implicated in nearly 50% of all cancers.

Leveraging CRISPR/Cas9 gene editing and optimized cell painting with near-infrared (NIR) dyes, the study overcomes traditional spectral overlap challenges, allowing researchers to confidently assess the effects of gene knockout and drug treatment on cell morphology.

In this application note, researchers used the Thermo Scientific™ CellInsight™ CX7 LZR Pro High-Content Screening Platform with an adapted cell painting assay to investigate the phenotypic consequences of p53 knockout in A549 lung carcinoma cells. Leveraging CRISPR/Cas9 gene editing and optimized cell painting with near-infrared (NIR) dyes, the study overcomes traditional spectral overlap challenges, allowing researchers to confidently assess the effects of gene knockout and drug treatment on cell morphology.

What You’ll Learn:

How to successfully knock out p53 using Invitrogen™ LentiArray™ Cas9 and Neon™ Transfection System

How to optimize cell painting assays using NIR reagents to reduce spectral interference in multiplexed imaging

How wild-type vs. p53 knockout A549 cells differ in response to cytotoxic compounds like etoposide, staurosporine, and cytochalasin D

How the CX7 LZR Pro platform supports precise phenotypic profiling through advanced imaging and channel separation

Why It Matters

This study showcases how pairing CRISPR-edited cell lines with a refined, high- resolution imaging assay can unlock deeper phenotypic insights that are critical for cancer biology research and drug development. With platform-specific dye modifications and precise high-content analysis, researchers gain greater confidence in their data— without the limitations of traditional fluorescence overlap.

Download the full application note to see how phenotypic changes in p53 knockout cells were quantified with clarity and confidence.

DOWNLOAD THIS APPLICATION NOTE

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