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Experimental RNAi

Experimental RNAi

RNA interference (RNAi) is a cellular mechanism that inhibits gene expression by suppressing its transcription or activating the RNA degradation process. The mechanism was discovered by Andrew Fire and Craig Mello in 1998 in plants. Today, it is observed in almost all eukaryotes, including protozoa, flies, nematodes, insects, parasites, and mammals. This precise cellular mechanism of gene silencing has been developed into a technique that provides an efficient way to identify and determine the...

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Related Experiment Video

Updated: Jun 17, 2025

Efficient Genome Editing of Mice by CRISPR Electroporation of Zygotes

Efficient Genome Editing of Mice by CRISPR Electroporation of Zygotes

Published on: December 16, 2022

A CRISPR/RfxCas13d-mediated strategy for efficient RNA knockdown in mouse embryonic development.

Lin Zhang¹, Shi-Meng Cao², Hao Wu²

¹State Key Laboratory of Cell Biology, Shanghai Key Laboratory of Molecular Andrology, Shanghai Institute of Biochemistry and Cell Biology, Center for Excellence in Molecular Cell Science, Chinese Academy of Sciences, University of Chinese Academy of Sciences, Shanghai, 200031, China.

Science China. Life Sciences

|August 7, 2024

Summary

This study introduces a novel method combining CRISPR-RfxCas13d and semi-cloning to suppress specific RNA levels in vivo. This technique effectively revealed critical roles of Sfmbt2 mRNA, Fendrr lncRNA, and circMan1a2 in embryonic development.

Keywords:

CRISPR-RfxCas13d RNA knockdown embryonic development semi-cloned technology sperm-like stem cell

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Last Updated: Jun 17, 2025

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Published on: April 2, 2020

Area of Science:

Molecular Biology
Developmental Biology
Genetics

Background:

Diverse RNA classes (mRNA, lncRNA, circRNA) are crucial for development and disease.
Limited understanding of RNA functions in vivo due to lack of effective RNA manipulation tools.

Purpose of the Study:

To develop and validate a novel system for targeted RNA suppression in live organisms.
To investigate the roles of specific RNAs (Sfmbt2 mRNA, Fendrr lncRNA, circMan1a2) in embryonic development.

Main Methods:

Utilized the CRISPR-RfxCas13d system for RNA targeting.
Employed sperm-like stem cell-mediated semi-cloning for gene delivery and expression suppression.
Interfered with the expression of Sfmbt2 mRNA, Fendrr lncRNA, and circMan1a2.

Main Results:

Successfully suppressed the expression of targeted mRNA, lncRNA, and circRNA molecules.
Demonstrated critical roles of these RNAs in embryonic development, evidenced by phenotypes like lethality and developmental delay.
Observed embryo resorption upon suppression of specific RNA targets.

Conclusions:

The developed CRISPR-RfxCas13d and semi-cloning method provides a powerful tool for targeted RNA silencing in vivo.
This methodology enables functional studies of specific RNAs without genetic modification.
The study highlights the essential roles of Sfmbt2 mRNA, Fendrr lncRNA, and circMan1a2 in successful embryonic development.