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Mouse embryonic stem cells as a model genetic system to dissect and exploit the RNA interference machinery
Stefan A Muljo1, Chryssa Kanellopoulou
1The CBR Institute for Biomedical Research and Department of Pathology, Harvard Medical School, Boston, MA, USA.
Methods in Molecular Biology (Clifton, N.J.)
|September 8, 2006
Summary
Conditional gene targeting in embryonic stem cells enables rapid investigation of RNA interference (RNAi) pathway functions in mammals. This method offers an economical and efficient alternative to traditional mouse models for gene function studies.
Area of Science:
- Genetics
- Molecular Biology
- Mammalian Systems
Background:
- Conditional gene targeting is crucial for understanding gene function in vivo.
- The RNA interference (RNAi) pathway plays a significant role in mammalian gene regulation.
- Embryonic stem (ES) cells offer a versatile platform for genetic manipulation in mammals.
Purpose of the Study:
- To investigate the biological role of the RNA interference (RNAi) pathway in mammals.
- To explore the utility of genetically engineered embryonic stem (ES) cells for gene function studies.
- To demonstrate the application of RNAi machinery in mammalian cells for gene knock-down experiments.
Main Methods:
- Utilizing conditional gene targeting in genetically engineered embryonic stem (ES) cells.
- Employing RNA interference (RNAi) for gene knock-down in mammalian cells.
- Performing mutagenesis and experimentation within a mammalian genetic system.
Main Results:
- Genetically engineered ES cells provide a physiological and tractable system for mutagenesis.
- The described approach allows for economical and rapid gene function elucidation.
- RNAi machinery in mammalian cells can be effectively exploited for gene knock-down.
Conclusions:
- Genetically engineered ES cells are a powerful tool for studying gene function, including the RNAi pathway, in mammals.
- This methodology offers a faster and more cost-effective alternative to generating genetically engineered mice.
- The chapter details experimental strategies using ES cells for genetic research.
Related Concept Videos
02:15Experimental 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...
01:23RNA Interference
RNA interference (RNAi) is a process in which a small non-coding RNA molecule blocks the post-transcriptional expression of a gene by binding to its messenger RNA (mRNA) and preventing the protein from being translated.
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...
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01:23RNA Interference
RNA interference (RNAi) is a process in which a small non-coding RNA molecule blocks the post-transcriptional expression of a gene by binding to its messenger RNA (mRNA) and preventing the protein from being translated.
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...
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...
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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.