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A Multiplexed Luciferase-based Screening Platform for Interrogating Cancer-associated Signal Transduction in Cultured Cells

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Identification of Target Genes Using Innovative Screening Systems.

Keisuke Sugita^1,2,3, Morito Kurata^1,4

¹Department of Comprehensive Pathology, Graduate School of Medical and Dental Sciences, Institute of Science Tokyo, Tokyo, Japan.

Pathology International

|May 6, 2025

Summary

Next-generation sequencing and CRISPR screening advance cancer research by identifying key genes for survival and drug resistance. These powerful genetic tools, including single-cell and spatial analyses, offer deeper insights into tumor complexity and therapeutic development.

Keywords:

CRISPR screening drug resistance oncogene random mutagenesis reverse genetic screening

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Area of Science:

Genomics
Cancer Biology
Genetic Engineering

Background:

Next-generation sequencing (NGS) has revolutionized cancer biology by identifying oncogenes and tumor suppressor genes.
Single-cell analysis and reverse genetic screenings are crucial for understanding cancer progression and heterogeneity.
CRISPR screening has become a powerful tool for functional genomics in cancer research due to its efficiency and simplicity.

Purpose of the Study:

To explore the role of gene function in cancer using advanced genetic screening techniques.
To investigate cancer progression, heterogeneity, and drug resistance mechanisms.
To highlight the advancements in CRISPR screening and its integration with single-cell and spatial analyses for in vivo mimicry.

Main Methods:

Utilizing reverse genetic screening methods, including random mutagenesis (retroviruses, transposons, RNA interference) and CRISPR.
Employing CRISPR screening and its modifications for comprehensive genetic mutation induction.
Integrating single-cell and spatial analyses with CRISPR screening to study cell-cell and spatial interactions.

Main Results:

Identification of essential genes involved in cancer survival, drug resistance, and tumorigenesis.
Enhanced understanding of cancer progression and heterogeneity through advanced screening techniques.
CRISPR screening, especially when combined with single-cell and spatial analyses, provides deeper insights into in vivo microenvironments.

Conclusions:

CRISPR screening and related technologies are significantly advancing cancer research.
These genetic tools are vital for uncovering complex biological processes in cancer.
The integration of various screening and analysis techniques accelerates the development of novel cancer therapeutics.