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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 005 01 - Diskussion
Zuletzt bearbeitet: 2014-10-28 05:11:46 Hindemith
Fragment, Gesichtet, SMWFragment, Schutzlevel sysop, Verschleierung, Wikipedia microRNA 2007, Ww

Typus
Verschleierung
Bearbeiter
SleepyHollow02
Gesichtet
Yes.png
Untersuchte Arbeit:
Seite: 5, Zeilen: 1-18
Quelle: Wikipedia microRNA 2007
Seite(n): 1 (Internetquelle), Zeilen: -
[In such cases, the formation of the double-stranded RNA through the binding of the miRNA triggers the degradation of the mRNA transcript through a process similar to RNAi, though] in other cases it is believed that the miRNA complex blocks the protein translation machinery or otherwise prevents protein translation without causing the mRNA to be degraded. miRNAs may also target methylation of genomic sites which correspond to targeted mRNAs. miRNAs function in association with a complement of proteins collectively termed the miRNP (micro-ribonucleic protein).

This effect was first described for the worm C. elegans in 1993 (Lee et al., 1993). As of 2002, miRNAs have been confirmed in various plants and animals, including C. elegans, human and the plant Arabidopsis thaliana. Genes have been found in bacteria that are similar in the sense that they control mRNA abundance or translation by binding a mRNA by base pairing, however they are not generally considered to be miRNAs because the Dicer enzyme is not involved.

In plants, similar RNA species termed short-interfering RNAs are used to prevent the transcription of viral RNA. While this siRNA is double-stranded, the mechanism seems to be closely related to that of miRNAs, especially taking the hairpin structures into account. siRNAs are also used to regulate cellular genes, as miRNAs do.

In such cases, the formation of the double-stranded RNA through the binding of the miRNA triggers the degradation of the mRNA transcript through a process similar to RNA interference (RNAi), though in other cases it is believed that the miRNA complex blocks the protein translation machinery or otherwise prevents protein translation without causing the mRNA to be degraded. miRNAs may also target methylation of genomic sites which correspond to targeted mRNAs. miRNAs function in association with a complement of proteins collectively termed the miRNP.

This effect was first described for the worm C. elegans in 1993 by Victor Ambros and coworkers (Lee et al., 1993). As of 2002, miRNAs have been confirmed in various plants and animals, including C. elegans, human and the plant Arabidopsis thaliana. Genes have been found in bacteria that are similar in the sense that they control mRNA abundance or translation by binding an mRNA by base pairing, however they are not generally considered to be miRNAs because the Dicer enzyme is not involved.

In plants, similar RNA species termed short-interfering RNAs siRNAs are used to prevent the transcription of viral RNA. While this siRNA is double-stranded, the mechanism seems to be closely related to that of miRNA, especially taking the hairpin structures into account. siRNAs are also used to regulate cellular genes, as miRNAs do.

Anmerkungen

Kein Hinweis auf die Quelle.

Sichter
(SleepyHollow02), Hindemith

[2.] Ww/Fragment 005 20 - Diskussion
Zuletzt bearbeitet: 2016-02-03 22:18:56 Schumann
Fragment, Gesichtet, SMWFragment, Schutzlevel sysop, Verschleierung, Wikipedia Small hairpin RNA 2007, Ww

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Hindemith
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Untersuchte Arbeit:
Seite: 5, Zeilen: 20-24
Quelle: Wikipedia Small hairpin RNA 2007
Seite(n): 1 (Internetquelle), Zeilen: -
Short hairpin RNAs (shRNAs) are a sequence of RNAs that makes a tight hairpin turn and are transcribed by RNA polymerase Ⅲ. It uses a vector introduced into cells and utilizes the U6 promoter to ensure that the shRNA is always expressed and can be cleaved by the cellular machinery into siRNA, which is then bound to the RISC silencing gene expression via RNAi (Brummelkamp et al., 2002; Lee et al., 2002; Miyagishi and Tiara, 2002; [Paddison, et al. 2002; Paul et al., 2002; Sui et al., 2002; Cao et al, 2005; Harper et al., 2005; McIntyre and Fanning, 2006).] A short hairpin RNA (shRNA) is a sequence of RNA that makes a tight hairpin turn that can be used to silence gene expression via RNA interference. shRNA uses a vector introduced into cells and utilizes the U6 promoter to ensure that the shRNA is always expressed. This vector is usually passed on to daughter cells, allowing the gene silencing to be inherited. The shRNA hairpin structure is cleaved by the cellular machinery into siRNA, which is then bound to the RNA-induced silencing complex (RISC).

[...]

References

McIntyre G, Fanning G (2006). "Design and cloning strategies for constructing shRNA expression vectors". BMC Biotechnol. 6: 1. PMID 16396676.

Harper S, Staber P, He X, Eliason S, Martins I, Mao Q, Yang L, Kotin R, Paulson H, Davidson B (2005). "RNA interference improves motor and neuropathological abnormalities in a Huntington's disease mouse model". Proc. Natl. Acad. Sci. U.S.A. 102 (16): 5820–5. PMID 15811941.

Nielsen M, Pedersen F, Kjems J (2005). "Molecular strategies to inhibit HIV-1 replication". Retrovirology 2: 10. PMID 15715913.

Paddison P, Caudy A, Bernstein E, Hannon G, Conklin D (2002). "Short hairpin RNAs (shRNAs) induce sequence-specific silencing in mammalian cells". Genes Dev. 16 (8): 948–58. PMID 11959843.

Cao W, Hunter R, Strnatka D, McQueen C, Erickson R (2005). "DNA constructs designed to produce short hairpin, interfering RNAs in transgenic mice sometimes show early lethality and an interferon response". J. Appl. Genet. 46 (2): 217–25. PMID 15876690.

Anmerkungen

Ein Verweis auf die Quelle fehlt.

Sichter
(Hindemith) Schumann


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Letzte Bearbeitung dieser Seite: durch Benutzer:Schumann, Zeitstempel: 20160203222037

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