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In vitro Mutagenesis

In vitro Mutagenesis

To learn more about the function of a gene, researchers can observe what happens when the gene is inactivated or “knocked out,” by creating genetically engineered knockout animals. Knockout mice have been particularly useful as models for human diseases such as cancer, Parkinson’s disease, and diabetes.

In-vitro Mutagenesis

In-vitro Mutagenesis

To learn more about the function of a gene, researchers can observe what happens when the gene is inactivated or “knocked out,” by creating genetically engineered knockout animals. Knockout mice have been particularly useful as models for human diseases such as cancer, Parkinson’s disease, and diabetes.

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Related Experiment Video

Updated: Jun 2, 2026

In Vitro Cleavage Assays using Purified Recombinant Drosophila Caspases for Substrate Screening

In Vitro Cleavage Assays using Purified Recombinant Drosophila Caspases for Substrate Screening

Published on: October 6, 2022

In vitro RISC cleavage assay.

Julia Stoehr¹, Gunter Meister

¹Center for Integrated Protein Science Munich (CIPSM), Laboratory for RNA Biology, Max-Planck-Institute of Biochemistry, Martinsried, Germany.

Methods in Molecular Biology (Clifton, N.J.)

|April 30, 2011

Summary

This summary is machine-generated.

RNA interference (RNAi) utilizes short-interfering RNAs (siRNAs) to guide the RNA-induced silencing complex (RISC) for target RNA cleavage. This chapter details a protocol for in vitro RISC assays to study gene expression.

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Last Updated: Jun 2, 2026

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Published on: May 3, 2014

Area of Science:

Molecular Biology
Genetics

Background:

RNA interference (RNAi) is a crucial biological process for gene silencing.
Short-interfering RNAs (siRNAs) are key mediators in the RNAi pathway.
The RNA-induced silencing complex (RISC) is responsible for target RNA recognition and cleavage.

Purpose of the Study:

To provide a detailed protocol for performing in vitro RNA-induced silencing complex (RISC) assays.
To facilitate the characterization of the RNA interference pathway.
To support the analysis of gene expression using RNAi.

Main Methods:

Detailed step-by-step protocol for in vitro RISC assays.
Description of necessary reagents and equipment.
Guidance on assay optimization and data interpretation.

Main Results:

The protocol enables the study of RISC loading and target binding.
The assays allow for the direct observation of RNA cleavage activity.
Successful application of the protocol across various organisms is implied.

Conclusions:

In vitro RISC assays are powerful tools for understanding RNAi mechanisms.
This protocol offers a standardized method for researchers studying gene silencing.
The methodology aids in the functional characterization of RNAi components.