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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 007 01 - Diskussion
Zuletzt bearbeitet: 2014-10-29 17:07:10 Hindemith
Fragment, Gesichtet, KomplettPlagiat, SMWFragment, Schutzlevel sysop, Wikipedia Small hairpin RNA 2007, Ww

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Quelle: Wikipedia Small hairpin RNA 2007
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shRNAs can also be made for use in plants and other systems, and are not necessarily driven by a U6 promoter. In plants the traditional promoter for strong constitutive [sic] expression (in most plant species) is the cauliflower mosaic virus 35S promoter (CaMV35S), in which case RNA polymerase II is used to express the transcript destined to initiate RNAi. shRNAs can also be made for use in plants and other systems, and are not necessarily driven by a U6 promoter. In plants the traditional promoter for strong consitutive expression (in most plant species) is the cauliflower mosaic virus 35S promoter (CaMV35S), in which case RNA Polymerase II is used to express the transcript destined to initiate RNAi
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[2.] Ww/Fragment 007 07 - Diskussion
Zuletzt bearbeitet: 2014-10-29 17:07:06 Hindemith
Fragment, Gesichtet, SMWFragment, Schutzlevel sysop, Verschleierung, Ww, Zou and Yoder 2005

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Quelle: Zou and Yoder 2005
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RNA interference (RNAi) was a term coined by Fire and coworkers to describe the inhibition of gene expression by double-stranded RNAs (dsRNAs) when introduced into nematode worms (Caenorhabditis elegans). Following on from the studies of Guo and Kemphues (1995), who had found that sense RNA was as effective as antisense RNA for suppressing gene expression in worms, Fire et al. (1998) applied single-stranded antisense RNA and double stranded RNA in their experiments. To their surprise, they found that dsRNA was more effective in producing interference than was either strand individually. After injection into adult C. elegans, single-stranded antisense RNA had a modest effect in diminishing specific gene expression, whereas double-stranded mixtures caused potent and specific interference.

Today we know that RNAi is a multistep process involving the generation of small interfering RNAs (siRNAs) in vivo through the action of the RNase III endonuclease “Dicer”. The resulting 21- to 23-nt siRNAs mediate the degradation of their complementary RNA (Shi, 2003).

RNA interference (RNAi) was a term coined by Fire and coworkers (1998) to describe the inhibition of gene expression by double-stranded RNAs (dsRNAs) when introduced into nematode worms (Caenorhabditis elegans). Following on from the studies of Guo and Kemphues (1995), who had found that sense RNA was as effective as antisense RNA for suppressing gene expression in worms, Fire et al. (1998) applied single-stranded antisense RNA and double stranded RNA in their experiments. To their surprise, they found that dsRNA was more effective at producing interference than was either strand individually. After injection into adult C. elegans, single-stranded antisense RNA had a modest effect in diminishing specific gene expression, whereas double-stranded mixtures caused potent and specific interference (Fire et al., 1998). RNAi is a multistep process involving the generation of small interfering RNAs (siRNAs) in vivo through the action of the RNase III endonuclease ‘Dicer’. The resulting 21- to 23-nt siRNAs mediate degradation of their complementary RNA (Shi, 2003).
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[3.] Ww/Fragment 007 22 - Diskussion
Zuletzt bearbeitet: 2014-10-29 17:07:02 Hindemith
Fragment, Geley and Mueller 2004, Gesichtet, KomplettPlagiat, SMWFragment, Schutzlevel sysop, Ww

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A major breakthrough in the elucidation of the underlying mechanism was the biochemical analysis of RNAi using Drosophila embryo or cell extracts (Tuschl et al., 1999; Hammond et al., 2000; Zamore et al., 2000), which led to the identification of the dsRNA processing enzyme Dicer (Bernstein et al., 2001a) as well as the RNA induced silencing complex, RISC (Hammond et al., 2000), which executes RNAi by using the small dsRNA species generated by Dicer as guidance molecules to target the homologous, endogenous mRNA for [degradation (Elbashir et al., 2001b,c; Zamore et al., 2000).] A major breakthrough in the elucidation of the underlying mechanism was the biochemical analysis of RNAi using Drosophila embryo or cell extracts (Hammond et al., 2000; Tuschl et al., 1999; Zamore et al., 2000), which led to the identification of the dsRNA processing enzyme Dicer (Bernstein et al., 2001a) as well as the RNA induced silencing complex, RISC (Hammond et al., 2000), which executes RNAi by using the small dsRNA species generated by Dicer as guidance molecules to target the homologous, endogenous mRNA for degradation (Elbashir et al., 2001b,c; Zamore et al., 2000).
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