Genome-wide resources of endoribonuclease-prepared short interfering RNAs for specific loss-of-function studies.
RNA interference (RNAi) has become an important technique for loss-of-gene-function studies in mammalian cells. To achieve reliable results in an RNAi experiment, efficient and specific silencing triggers are required. Here we present genome-wide data sets for the production of endoribonuclease-prepared short interfering RNAs (esiRNAs) for human, mouse and rat. We used an algorithm to predict the optimal region for esiRNA synthesis for every protein-coding gene of these three species. We created a database, RiDDLE, for retrieval of target sequences and primer information. To test this in silico resource experimentally, we generated 16,242 esiRNAs that can be used for RNAi screening in human cells. Comparative analyses with chemically synthesized siRNAs demonstrated a high silencing efficacy of esiRNAs and a 12-fold reduction of downregulated off-target transcripts as detected by microarray analysis. Hence, the presented esiRNA libraries offer an efficient, cost-effective and specific alternative to presently available mammalian RNAi resources.
Kittler
Ralf
R
Max Planck Institute for Molecular Cell Biology and Genetics, Pfotenhauerstrasse 108, D-01307 Dresden, Germany.
Surendranath
Vineeth
V
Heninger
Anne-Kristin
AK
Slabicki
Mikolaj
M
Theis
Mirko
M
Putz
Gabriele
G
Franke
Kristin
K
Caldarelli
Antonio
A
Grabner
Hannes
H
Kozak
Karol
K
Wagner
Jan
J
Rees
Effi
E
Korn
Bernd
B
Frenzel
Corina
C
Sachse
Christoph
C
Sönnichsen
Birte
B
Guo
Jie
J
Schelter
Janell
J
Burchard
Julja
J
Linsley
Peter S
PS
Jackson
Aimee L
AL
Habermann
Bianca
B
Buchholz
Frank
F
D000818:Animals; D004722:Endoribonucleases / Q000235:genetics*; D015698:Genomic Library; D023281:Genomics / Q000379:methods*; D006801:Humans; D034622:RNA Interference; D034741:RNA, Small Interfering / Q000235:genetics*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012680:Sensitivity and Specificity; D014158:Transcription, Genetic; D014162:Transfection; D020506:Untranslated Regions; D014584:User-Computer Interface
D034741:RNA, Small Interfering; D020506:Untranslated Regions; D004722:Endoribonucleases