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Validation of shRNA clones for gene silencing in 293FT cells

von Dr. Wen Wang

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[1.] Ww/Fragment 002 01 - Diskussion
Zuletzt bearbeitet: 2014-10-28 02:51:17 Hindemith
Fragment, Gesichtet, SMWFragment, Schutzlevel sysop, Verschleierung, Wikipedia Small interfering RNA 2007, Ww

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Quelle: Wikipedia Small interfering RNA 2007
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[Most notably, siRNAs are involved in the RNA interference (RNAi) pathway] where the siRNA interferes with the expression of a specific gene (Tuschl et al, 2001). In addition to their role in the RNAi pathway, siRNAs also act in RNAi-related pathways, as an antiviral mechanism or in shaping the chromatin structure of a genome. siRNAs can also be exogenously (artificially) introduced into cells by various transfection methods to bring about the specific knockdown of a gene of interest. Essentially any gene of which the sequence is known can thus be targeted based on sequence complementarity with tailored siRNA. This has made siRNA an important tool for gene function and drug target validation studies in the post-genomic era. Most notably, siRNA is involved in the RNA interference (RNAi) pathway where the siRNA interferes with the expression of a specific gene. In addition to their role in the RNAi pathway, siRNAs also act in RNAi-related pathways, e.g. as an antiviral mechanism or in shaping the chromatin structure of a genome; the complexity of these pathways is only now being elucidated.

[...]

[...] SiRNAs can also be exogenously (artificially) introduced into cells by various transfection methods to bring about the specific knockdown of a gene of interest. Essentially any gene of which the sequence is known can thus be targeted based on sequence complementarity with an appropriately tailored siRNA. This has made siRNAs an important tool for gene function and drug target validation studies in the post-genomic era.

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[2.] Ww/Fragment 002 17 - Diskussion
Zuletzt bearbeitet: 2014-10-28 05:04:15 Hindemith
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In genetics, miRNAs are single-stranded RNA molecules of about 21-23 nucleotides in length, which regulate gene expression. miRNAs are encoded by genes that are transcribed from DNA but not translated into protein (non-coding RNA); instead they are processed from primary transcripts known as pri-miRNAs to short stem-loop structures called pre-miRNAs and finally to function miRNAs. Mature miRNA molecules are partially complementary to one or more messenger RNA (mRNA) molecules, and their main function is to down-regulate gene expression.

1.1.3.1 Formation and processing of miRNA

The genes encoding miRNAs are much longer than the processed mature miRNA molecule; miRNAs are first transcribed as primary transcripts or pri-miRNAs with a cap and poly-A tail and processed to short, 70-nucleotide stem-loop structures known as pre-miRNAs in the cell nucleus. This [processing is performed in animals by a protein complex known as the Microprocessor complex, consisting of the nuclease Drosha and the and the double-stranded RNA binding protein Pasha (Denli et al, 2004).]

In genetics, microRNAs (miRNA) are single-stranded RNA molecules of about 21-23 nucleotides in length regulating gene expression. miRNAs are encoded by genes that are transcribed from DNA but not translated into protein (non-coding RNA); instead they are processed from primary transcripts known as pri-miRNA to short stem-loop structures called pre-miRNA and finally to functional miRNA. Mature miRNA molecules are partially complementary to one or more messenger RNA (mRNA) molecules, and their main function is to downregulate gene expression. [...]

[...]

The genes encoding miRNAs are much longer than the processed mature miRNA molecule; miRNAs are first transcribed as primary transcripts or pri-miRNA with a cap and poly-A tail and processed to short, 70-nucleotide stem-loop structures known as pre-miRNA in the cell nucleus. This processing is performed in animals by a protein complex known as the Microprocessor complex, consisting of the nuclease Drosha and the double-stranded RNA binding protein Pasha.[3]


3. Denli AM, Tops BB, Plasterk RH, Ketting RF, Hannon GJ. (2004). Nature 432(7014):231-5.

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