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Updated: Jul 5, 2026
Biotin-based Pulldown Assay to Validate mRNA Targets of Cellular miRNAs
Published on: June 12, 2018
Target validation to biomarker development: focus on RNA interference
Riccardo Colombo1, Jürgen Moll
1Nerviano Medical Sciences Srl, Nerviano, Italy.
Abstract:
With the growing number of putative molecular targets and increased economic pressure on companies developing novel drugs, particularly in the cancer area, the need to work on highly validated targets is essential. The use of biomarkers for proof of mechanism of action is becoming an important tool in validation efforts in the preclinical phase of drug development, helping to reduce the attrition rate of candidate drugs once they have entered the clinic. In this review, we highlight how RNA interference (RNAi) has become the method of choice to perform both target validation and identification in academia and industry. RNAi takes advantage of a naturally occurring mechanism whereby cells regulate the expression of genes at the post-transcriptional level, and it introduces a new era in loss-of-function experiments, allowing for the rapid measurement of the phenotype observed upon target expression abrogation. Design of both small-interfering RNA and short-hairpin RNA constructs and their delivery into cells have emerged as the most important aspects of this technology, and reduction or measurement of potential unwanted off-target effects must also be taken into consideration. A number of successes have already been described, and several oncology targets and biomarkers have been identified and validated with this technique.
Insights
RNA interference (RNAi) is crucial for validating cancer drug targets. This method enables rapid loss-of-function studies, identifying and validating oncology targets and biomarkers effectively.
Area of Science:
- Molecular Biology
- Drug Discovery
- Oncology
Background:
- Increasingly, the pharmaceutical industry faces economic pressure to validate molecular targets, especially in cancer drug development.
- Biomarkers are vital for proving mechanism of action in preclinical drug development, reducing clinical attrition rates.
Purpose of the Study:
- To review the role of RNA interference (RNAi) as a key technology for target validation and identification.
- To highlight the application of RNAi in academic and industrial settings for preclinical drug development.
Main Methods:
- RNA interference (RNAi) leverages a natural cellular mechanism for post-transcriptional gene regulation.
- Small-interfering RNA (siRNA) and short-hairpin RNA (shRNA) design and delivery are critical components.
- Minimizing off-target effects is essential for accurate validation studies.
Main Results:
- RNAi facilitates rapid loss-of-function experiments to observe phenotypes upon target gene abrogation.
- The technique has demonstrated success in identifying and validating numerous oncology targets.
- RNAi has proven effective in discovering and validating cancer biomarkers.
Conclusions:
- RNA interference (RNAi) is the method of choice for target validation and identification in drug discovery.
- This technology significantly aids in reducing drug candidate attrition by validating targets early in development.
- Successful applications in oncology demonstrate the power of RNAi for identifying novel therapeutic targets and biomarkers.
Related Concept Videos
02:15Experimental RNAi
01:23RNA Interference
This process occurs naturally in cells, often through the activity of genomically-encoded microRNAs. Researchers can take advantage of this mechanism by introducing synthetic RNAs to deactivate specific genes for research or therapeutic purposes. For example, RNAi could be used...
01:23RNA Interference
This process occurs naturally in cells, often through the activity of genomically-encoded microRNAs. Researchers can take advantage of this mechanism by introducing synthetic RNAs to deactivate specific genes for research or therapeutic purposes. For example, RNAi could be used...
01:22MicroRNAs