RNAi induction and activation in mammalian muscle cells where Dicer and eIF2C translation initiation factors are barely expressed.
Dicer plays an important role in the course of RNA interference (RNAi), i.e., it digests long double-stranded RNAs into 21-25 nucleotide small-interfering RNA (siRNA) duplexes functioning as sequence-specific RNAi mediators. In this study, we investigated the expression levels of Dicer and eIF2C1 approximately 4, which, like Dicer, appear to participate in mammalian RNAi, in various mouse tissues. Results indicate that the levels of eIF2C1 approximately 4 as well as Dicer are lower in skeletal muscle and heart than in other tissues. To see if RNAi could occur under such a condition with low levels of expression of Dicer and eIF2C1 approximately 4, we examined RNAi activity in mouse skeletal muscle fibers. The results indicate that RNAi can be induced by synthetic siRNA duplexes in muscle fibers. We further examined RNAi activity during myogenic differentiation of mouse C2C12 cells. The data indicate that although the expression levels of Dicer and eIF2C1 approximately 4 decrease during the differentiation, RNAi can be induced in the cells. Altogether, the data presented here suggest that muscle cells retain the ability to induce RNAi, although Dicer and eIF2C1 approximately 4 appear to be barely expressed in them.
Sago
Noriko
N
National Institute of Neuroscience, NCNP, 4-1-1 Ogawahigashi, Kodaira, Tokyo 187-8502, Japan.
Omi
Kazuya
K
Tamura
Yoshiko
Y
Kunugi
Hiroshi
H
Toyo-oka
Teruhiko
T
Tokunaga
Katsushi
K
Hohjoh
Hirohiko
H
D000818:Animals; D060565:Argonaute Proteins; D001973:Bromodeoxyuridine / Q000494:pharmacology; D002453:Cell Cycle; D002454:Cell Differentiation; D002460:Cell Line; D004396:Coloring Agents / Q000494:pharmacology; D053487:DEAD-box RNA Helicases; D018076:DNA, Complementary / Q000378:metabolism; D004305:Dose-Response Relationship, Drug; D004722:Endoribonucleases / Q000096:biosynthesis*; D039642:Eukaryotic Initiation Factors / Q000096:biosynthesis*; D017930:Genes, Reporter; D008156:Luciferases / Q000378:metabolism; D051379:Mice; D018482:Muscle, Skeletal / Q000378:metabolism; D009132:Muscles / Q000378:metabolism*; D009841:Oligonucleotides / Q000737:chemistry; D014176:Protein Biosynthesis; D012313:RNA / Q000378:metabolism; D020365:RNA Helicases / Q000096:biosynthesis*; D034622:RNA Interference; D020133:Reverse Transcriptase Polymerase Chain Reaction; D043244:Ribonuclease III; D013997:Time Factors; D014018:Tissue Distribution; D014162:Transfection
C115848:AGO1 protein, human; D060565:Argonaute Proteins; D004396:Coloring Agents; D018076:DNA, Complementary; D039642:Eukaryotic Initiation Factors; D009841:Oligonucleotides; D012313:RNA; D008156:Luciferases; D004722:Endoribonucleases; C408087:DICER1 protein, human; D043244:Ribonuclease III; D053487:DEAD-box RNA Helicases; D020365:RNA Helicases; D001973:Bromodeoxyuridine