Antisense-mediated depletion reveals essential and specific functions of microRNAs in Drosophila development.
MicroRNAs are small noncoding RNAs that control gene function posttranscriptionally through mRNA degradation or translational inhibition. Much has been learned about the processing and mechanism of action of microRNAs, but little is known about their biological function. Here, we demonstrate that injection of 2'O-methyl antisense oligoribonucleotides into early Drosophila embryos leads to specific and efficient depletion of microRNAs and thus permits systematic loss-of-function analysis in vivo. Twenty-five of the forty-six embryonically expressed microRNAs show readily discernible defects; pleiotropy is moderate and family members display similar yet distinct phenotypes. Processes under microRNA regulation include cellularization and patterning in the blastoderm, morphogenesis, and cell survival. The largest microRNA family in Drosophila (miR-2/6/11/13/308) is required for suppressing embryonic apoptosis; this is achieved by differential posttranscriptional repression of the proapoptotic factors hid, grim, reaper, and sickle. Our findings demonstrate that microRNAs act as specific and essential regulators in a wide range of developmental processes.
Laboratory of Developmental Neurogenetics, Rockefeller University, 1230 York Ave, New York, New York 10021, USA.
Dan
D
Leaman
Po Yu
PY
Chen
John
J
Fak
Abdullah
A
Yalcin
Michael
M
Pearce
Ulrich
U
Unnerstall
Debora S
DS
Marks
Chris
C
Sander
Thomas
T
Tuschl
Ulrike
U
Gaul
D000818:Animals; D017209:Apoptosis / Q000235:genetics; D019521:Body Patterning / Q000187:drug effects / Q000235:genetics*; D002470:Cell Survival / Q000235:genetics; D015536:Down-Regulation / Q000187:drug effects / Q000235:genetics; D029721:Drosophila Proteins / Q000235:genetics; D004331:Drosophila melanogaster / Q000196:embryology* / Q000235:genetics* / Q000378:metabolism; D004625:Embryo, Nonmammalian / Q000196:embryology / Q000378:metabolism; D047108:Embryonic Development / Q000187:drug effects / Q000235:genetics*; D018507:Gene Expression Regulation, Developmental / Q000187:drug effects / Q000235:genetics*; D035683:MicroRNAs / Q000037:antagonists & inhibitors / Q000235:genetics*; D009479:Neuropeptides / Q000235:genetics; D020319:Oligodeoxyribonucleotides, Antisense / Q000494:pharmacology; D012323:RNA Processing, Post-Transcriptional / Q000235:genetics*; D012097:Repressor Proteins / Q000235:genetics / Q000378:metabolism
D029721:Drosophila Proteins; C108880:HID protein, Drosophila; D035683:MicroRNAs; D009479:Neuropeptides; D020319:Oligodeoxyribonucleotides, Antisense; D012097:Repressor Proteins; C108881:grim protein, Drosophila; C086973:rpr protein, Drosophila; C452430:skl protein, Drosophila