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Updated: Feb 28, 2026
Dual CRISPR-Interference Strategy for Targeting Synthetic Lethal Interactions Between Non-Coding RNAs in Cancer Cells
Published on: May 30, 2025
State of the art technologies to explore long non-coding RNAs in cancer
Saeede Salehi1,2,3, Mohammad Naser Taheri1,2,3, Negar Azarpira4
1Diagnostic Laboratory Sciences and Technology Research Center, School of Paramedical Sciences, Shiraz University of Medical Sciences, Shiraz, Iran.
Abstract:
Long non-coding RNAs (lncRNAs) comprise a vast repertoire of RNAs playing a wide variety of crucial roles in tissue physiology in a cell-specific manner. Despite being engaged in myriads of regulatory mechanisms, many lncRNAs have still remained to be assigned any functions. A constellation of experimental techniques including single-molecule RNA in situ hybridization (sm-RNA FISH), cross-linking and immunoprecipitation (CLIP), RNA interference (RNAi), Clustered regularly interspaced short palindromic repeats (CRISPR) and so forth has been employed to shed light on lncRNA cellular localization, structure, interaction networks and functions. Here, we review these and other experimental approaches in common use for identification and characterization of lncRNAs, particularly those involved in different types of cancer, with focus on merits and demerits of each technique.
Insights
This review explores experimental methods for identifying and characterizing long non-coding RNAs (lncRNAs), focusing on their roles in cancer. It details techniques like sm-RNA FISH and CRISPR, evaluating their strengths and weaknesses.
Area of Science:
- Molecular Biology
- Genomics
- Cancer Research
Background:
- Long non-coding RNAs (lncRNAs) are crucial regulators of cell-specific tissue physiology.
- Many lncRNAs have unknown functions despite their diverse regulatory roles.
- Understanding lncRNA function is vital, especially in cancer biology.
Purpose of the Study:
- To review common experimental techniques for lncRNA identification and characterization.
- To focus on methods applicable to lncRNAs involved in various cancers.
- To critically assess the merits and demerits of each technique.
Main Methods:
- Single-molecule RNA in situ hybridization (sm-RNA FISH)
- Cross-linking and immunoprecipitation (CLIP)
- RNA interference (RNAi)
- Clustered regularly interspaced short palindromic repeats (CRISPR)
Main Results:
- The review details multiple experimental approaches for lncRNA analysis.
- Each method's advantages and disadvantages are discussed in the context of lncRNA research.
- The application of these techniques to cancer-related lncRNAs is highlighted.
Conclusions:
- A comprehensive understanding of experimental techniques is essential for advancing lncRNA research.
- Choosing the appropriate method is critical for accurate lncRNA functional characterization.
- This review provides a guide for researchers investigating lncRNAs in cancer and other fields.
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