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Related Concept Videos

Experimental RNAi02:15

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...
In-vitro Mutagenesis01:16

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

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Lentiviral-mediated Knockdown During Ex Vivo Erythropoiesis of Human Hematopoietic Stem Cells
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Published on: July 16, 2011

Efficient gene knockdowns in human embryonic stem cells using lentiviral-based RNAi.

Asmin Tulpule1, George Q Daley

  • 1Harvard Medical School, Karp Family Research Building, Division of Hematology/Oncology, Children's Hospital Boston, Boston, MA, USA.

Methods in Molecular Biology (Clifton, N.J.)
|December 18, 2008
PubMed
Summary

Lentiviral RNA interference (RNAi) offers an efficient method for gene knockdown in human embryonic stem cells (hESCs). This approach overcomes limitations of traditional gene editing, enabling faster research into human development and disease.

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Area of Science:

  • Stem cell biology
  • Molecular biology
  • Genetics

Background:

  • Human embryonic stem cells (hESCs) are vital for studying human development and disease.
  • Traditional gene manipulation methods like homologous recombination (HR) are inefficient in hESCs due to low cloning efficiency.
  • Developing effective tools for gene function studies in hESCs is crucial.

Purpose of the Study:

  • To present lentiviral RNA interference (RNAi) as a rapid and specific method for gene knockdown in hESCs.
  • To outline the essential requirements for successful lentiviral RNAi application in hESCs.
  • To facilitate research on human development and disease using hESCs.

Main Methods:

  • Utilizing lentiviral vectors for the delivery of small interfering RNAs (siRNAs) into hESCs.
  • Implementing RNA interference (RNAi) to achieve gene knockdown.
  • Comparing lentiviral siRNA delivery with traditional homologous recombination (HR) methods.

Main Results:

  • Lentiviral delivery of siRNAs enables rapid and specific gene knockdown in hESCs.
  • This method achieves highly efficient and persistent gene silencing.
  • It overcomes the challenges associated with low cloning efficiency in hESCs.

Conclusions:

  • Lentiviral RNAi is a powerful and efficient tool for functional genomics in hESCs.
  • This technique accelerates the study of gene function in human development and disease.
  • Key requirements for successful lentiviral RNAi in hESCs are summarized.