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Experimental RNAi

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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 7, 2026

CRISPR-Cas9 Mediated Gene Deletion in Human Pluripotent Stem Cells Cultured Under Feeder-Free Conditions

CRISPR-Cas9 Mediated Gene Deletion in Human Pluripotent Stem Cells Cultured Under Feeder-Free Conditions

Published on: November 1, 2024

High-efficiency siRNA-based gene knockdown in human embryonic stem cells.

Yinghong Ma¹, Jianyu Jin, Chunsheng Dong

¹Yale Stem Cell Center, New Haven, Connecticut 06520, USA.

RNA (New York, N.Y.)

|October 28, 2010

Summary

We developed a cost-effective, high-efficiency method for delivering molecules into human stem cells. This technique enables significant gene knockdown, facilitating studies on stem cell differentiation.

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Lentiviral-mediated Knockdown During Ex Vivo Erythropoiesis of Human Hematopoietic Stem Cells

Lentiviral-mediated Knockdown During Ex Vivo Erythropoiesis of Human Hematopoietic Stem Cells

Published on: July 16, 2011

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Last Updated: Jun 7, 2026

CRISPR-Cas9 Mediated Gene Deletion in Human Pluripotent Stem Cells Cultured Under Feeder-Free Conditions

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Published on: November 1, 2024

CRISPR/Cas9-Mediated Highly Efficient Gene Targeting in Embryonic Stem Cells for Developing Gene-Manipulated Mouse Models

CRISPR/Cas9-Mediated Highly Efficient Gene Targeting in Embryonic Stem Cells for Developing Gene-Manipulated Mouse Models

Published on: August 24, 2022

Lentiviral-mediated Knockdown During Ex Vivo Erythropoiesis of Human Hematopoietic Stem Cells

Lentiviral-mediated Knockdown During Ex Vivo Erythropoiesis of Human Hematopoietic Stem Cells

Published on: July 16, 2011

Area of Science:

Stem cell biology
Molecular biology
Gene delivery technologies

Background:

Loss-of-function studies in human embryonic stem cells (hESCs) and induced pluripotent stem cells (iPSCs) are crucial for understanding pluripotency and differentiation.
Existing nonviral delivery methods for these cells often suffer from low efficiency, hindering research progress.

Purpose of the Study:

To develop a simple, cost-effective, and high-efficiency nonviral method for delivering plasmids and small interfering RNAs (siRNAs) into hESCs and iPSCs.
To demonstrate the utility of this method for functional gene knockdown and subsequent analysis of stem cell differentiation.

Main Methods:

A novel nonviral delivery system was optimized for introducing genetic material into hESCs and iPSCs.
Small interfering RNAs (siRNAs) targeting key stem cell factors, Oct4 and Lin28, were delivered using the developed method.
Quantitative assessment of gene expression reduction and observation of cellular morphology and staining patterns were performed.

Main Results:

The developed method achieved high-efficiency delivery of plasmids and siRNAs into hESCs and iPSCs.
Delivery of siRNAs resulted in >90% reduction in the expression of Oct4 and Lin28.
Oct4 knockdown led to observable changes in cell morphology and staining patterns, indicative of stem cell differentiation.

Conclusions:

A simple, cost-effective, and highly efficient nonviral delivery method for hESCs and iPSCs has been established.
This method enables robust gene knockdown, facilitating functional studies of gene function in human pluripotent stem cells.
The findings provide a valuable tool for advancing research in stem cell biology and differentiation.