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Updated: Jun 27, 2025
Dual CRISPR-Interference Strategy for Targeting Synthetic Lethal Interactions Between Non-Coding RNAs in Cancer Cells
Published on: May 30, 2025
CRISPR-Cas9 screening identifies INTS3 as an anti-apoptotic RNA-binding protein and therapeutic target for colorectal
Zhiwei Wang1, Cheng Zhang1, Jing Guo1
1School of Life Sciences, Zhengzhou University, 100 Kexue Road, Zhengzhou 450001, China.
Abstract:
Growing evidences indicate that RNA-binding proteins (RBPs) play critical roles in regulating the RNA splicing, polyadenylation, stability, localization, translation, and turnover. Abnormal expression of RBPs can promote tumorigenesis. Here, we performed a CRISPR screen using an RBP pooled CRISPR knockout library and identified 27 potential RBPs with role in supporting colorectal cancer (CRC) survival. We found that the deletion/depletion of INTS3 triggered apoptosis in CRC. The in vitro experiments and RNA sequencing revealed that INTS3 destabilized pro-apoptotic gene transcripts and contributed to the survival of CRC cells. INTS3 loss delayed CRC cells growth in vivo. Furthermore, delivery of DOTAP/cholesterol-mshINTS3 nanoparticles inhibited CRC tumor growth. Collectively, our work highlights the role of INTS3 in supporting CRC survival and provides several novel therapeutic targets for treatment.
Insights
RNA-binding proteins (RBPs) are crucial in cancer. This study identified Integrator Complex Subunit 3 (INTS3) as vital for colorectal cancer (CRC) cell survival by destabilizing tumor-suppressing genes.
Area of Science:
- Molecular Biology
- Cancer Research
- Genetics
Background:
- RNA-binding proteins (RBPs) regulate gene expression and are implicated in tumorigenesis.
- Aberrant RBP expression is linked to cancer development and progression.
Purpose of the Study:
- To identify RBPs that support colorectal cancer (CRC) cell survival using a CRISPR knockout screen.
- To elucidate the specific role of Integrator Complex Subunit 3 (INTS3) in CRC.
Main Methods:
- Conducted a pooled CRISPR knockout screen utilizing an RBP library to identify essential RBPs in CRC.
- Performed in vitro and in vivo experiments, including RNA sequencing, to analyze the function of INTS3.
- Investigated the therapeutic potential of targeting INTS3 using nanoparticle delivery systems.
Main Results:
- Identified 27 RBPs supporting CRC survival, with INTS3 deletion triggering apoptosis in CRC cells.
- Demonstrated that INTS3 destabilizes pro-apoptotic gene transcripts, promoting CRC cell survival.
- Observed delayed CRC cell growth in vivo upon INTS3 loss and inhibited tumor growth with mshINTS3 nanoparticles.
Conclusions:
- INTS3 plays a critical role in maintaining colorectal cancer cell survival.
- INTS3 represents a potential therapeutic target for CRC treatment.
- Targeting INTS3, for example, via nanoparticle delivery, shows promise for inhibiting CRC tumor growth.
