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KeineWertung
Bearbeiter
Hindemith
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Untersuchte Arbeit:
Seite: 66, Zeilen: 3-16
Quelle: Krueger 2007
Seite(n): 237, 241, 246, 247, Zeilen: 237: 9ff; 241: r.col: 3ff, 246: r.col: last paragraph; 247: l.col: 1ff
Target gene-specific characteristics such as the accessibility of the corresponding target sequences to the RNAi appear to have a significant influence on the knockdown efficacy, making certain targets easy or difficult to knock down by using siRNA. It has been reported that low-abundant genes are less susceptible to siRNA-mediated knockdown (Hu et al., 2004). However, recently, Krueger et al. (2007) tested several thousand siRNAs for target genes from various gene families and showed that target genes from high or moderate to low expression levels were silenced equally well. Moreover, it was revealed that the localization of siRNA target sites on the transcript and siRNA concentration were not responsible for low silencing capacity of siRNA (Krueger et al., 2007). The performance of siRNA showing moderate silencing of < 70% knockdown threshold was almost unaffected by increasing the siRNA concentration used for transfection, by changing the cell type, or by increasing or decreasing the time point of analysis. We have functionally tested several thousand siRNAs for target genes from various gene families including kinases, phosphatases, and cancer-related genes (e.g., genes involved in apoptosis and the cell cycle). Some targets were difficult to silence above a threshold of 70% knockdown. [...] Target gene-specific characteristics such as the accessibility of the corresponding target sequences to the RNAi machinery appear to have a significant influence on the knockdown observed, making certain targets easy or difficult to knock down using siRNA.

[page 241]

However, it has been reported that low-abundant genes are less susceptible to siRNA-mediated knockdown (Hu et al., 2004). [...] This result shows that target genes with either high or moderate to low gene expression levels were silenced equally well. Therefore, target gene expression level does not serve as an indicator of successful silencing potential.

[page 246]

The performance of siRNAs showing moderate silencing of < 70% was almost unaffected by increasing the siRNA concentration used for transfection, by changing the cell type, or by increasing or decreasing the time

[page 247]

point of analysis.

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