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Hindemith
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Untersuchte Arbeit:
Seite: 11, Zeilen: 11-24
Quelle: Geley and Mueller 2004
Seite(n): 987, Zeilen: l.col: 1ff
1.2.1.2 Execution phase: assembly of siRNA containing silencing complexes

Dicer-generated siRNAs are then incorporated into a large multiprotein complex, which is involved in various gene-silencing modes, and is called the RNA induced silencing complex, or RISC (Hammond et al, 2000; Nykanen et al, 2001). Processing of dsRNA and assembly of a functional RISC likely occurs in the cytoplasm, as Dicer is a cytosolic enzyme and RISC activity can be purified from the cytosol (Billy et al, 2001). R2D2, a Drosophila gene related to the C. elegans RNAi gene RDE-4, has been implicated in the transfer of siRNAs into the RISC (Liu et al, 2003). Generation of siRNAs from dsRNA in Drosophila embryo extracts, unwinding of the siRNA duplex, and incorporation into the RISC requires ATP (Nykanen et al, 2001). In contrast, human Dicer does not seem to rely on ATP for processing of dsRNA into siRNA molecules (Zhang et al., 2002).

2.2. The execution phase: assembly of siRNA containing silencing complexes

Dicer-generated siRNAs are then incorporated into a large multiprotein complex, which is involved in various gene-silencing modes, and is called the RNA induced silencing complex, or RISC (Hammond et al., 2000; Nykanen et al., 2001). Processing of dsRNA and assembly of a functional RISC likely occurs in the cytoplasm, as Dicer is a cytosolic enzyme and RISC activity can be purified from cytosol (Billy et al., 2001). R2D2, a Drosophila gene related to the C. elegans RNAi gene RDE-4, has been implicated in the transfer of siRNAs into the RISC (Liu et al., 2003). Generation of siRNAs from dsRNA in Drosophila embryo extracts, unwinding of the siRNA duplex, and incorporation into the RISC require ATP (Nykanen et al., 2001). In contrast, human Dicer does not seem to require ATP for processing of dsRNA into siRNA molecules (Zhang et al., 2002).

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