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Vol. 13, Issue 2, 445-453, February 2002
and
*Abt. Genetik, Universität Kassel, D-34132 Kassel, Germany;
and We show that in Dictyostelium discoideum an
endogenous gene as well as a transgene can be silenced by introduction
of a gene construct that is transcribed into a hairpin RNA. Gene
silencing was accompanied by the appearance of sequence-specific RNA
~23mers and seemed to have a limited capacity. The three
Dictyostelium homologues of the RNA-directed RNA
polymerase (RrpA, RrpB, and DosA) all contain an N-terminal
helicase domain homologous to the one in the dicer nuclease, suggesting
exon shuffling between RNA-directed RNA polymerase and the dicer
homologue. Only the knock-out of rrpA resulted in a loss of the hairpin
RNA effect and simultaneously in a loss of detectable ~23mers.
However, ~23mers were still generated by the
Dictyostelium dsRNase in vitro with extracts from
rrpADepartment of Biochemistry and Molecular Biology, The
University of Chicago, Chicago, Illinois 60637-1432
, rrpB, and DosA cells.
Both RrpA and a target gene were required for production of detectable
amounts of ~23mers, suggesting that target sequences are involved in
~23mer amplification.
Corresponding author. E-mail address:
nellen{at}hrz.uni-kassel.de.
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