title: Depleting gene activities in early Drosophila embryos with the "maternal-Gal4-shRNA" system.
abstract: In a developing Drosophila melanogaster embryo, mRNAs have a maternal origin, a zygotic origin, or both. During the maternal-zygotic transition, maternal products are degraded and gene expression comes under the control of the zygotic genome. To interrogate the function of mRNAs that are both maternally and zygotically expressed, it is common to examine the embryonic phenotypes derived from female germline mosaics. Recently, the development of RNAi vectors based on short hairpin RNAs (shRNAs) effective during oogenesis has provided an alternative to producing germline clones. Here, we evaluate the efficacies of: (1) maternally loaded shRNAs to knockdown zygotic transcripts and (2) maternally loaded Gal4 protein to drive zygotic shRNA expression. We show that, while Gal4-driven shRNAs in the female germline very effectively generate phenotypes for genes expressed maternally, maternally loaded shRNAs are not very effective at generating phenotypes for early zygotic genes. However, maternally loaded Gal4 protein is very efficient at generating phenotypes for zygotic genes expressed during mid-embryogenesis. We apply this powerful and simple method to unravel the embryonic functions of a number of pleiotropic genes.
authors.affiliation: Department of Systems Biology, Harvard Medical School, Boston, MA 02115, USA.
authors.forename: Max V
authors.initials: MV
authors.lastname: Staller
authors.forename: Dong
authors.initials: D
authors.lastname: Yan
authors.forename: Sakara
authors.initials: S
authors.lastname: Randklev
authors.forename: Meghan D
authors.initials: MD
authors.lastname: Bragdon
authors.forename: Zeba B
authors.initials: ZB
authors.lastname: Wunderlich
authors.forename: Rong
authors.initials: R
authors.lastname: Tao
authors.forename: Lizabeth A
authors.initials: LA
authors.lastname: Perkins
authors.forename: Angela H
authors.initials: AH
authors.lastname: Depace
authors.forename: Norbert
authors.initials: N
authors.lastname: Perrimon
mesh_terms: D000818:Animals; D029721:Drosophila Proteins / Q000235:genetics* / Q000378:metabolism; D004331:Drosophila melanogaster / Q000196:embryology* / Q000235:genetics*; D005260:Female; D018628:Gene Dosage; D018507:Gene Expression Regulation, Developmental; D055785:Gene Knockdown Techniques; D008297:Male; D009866:Oogenesis / Q000235:genetics; D010641:Phenotype; D034622:RNA Interference; D034741:RNA, Small Interfering / Q000235:genetics* / Q000378:metabolism; D014157:Transcription Factors / Q000235:genetics* / Q000378:metabolism; D014158:Transcription, Genetic
chemical_list: D029721:Drosophila Proteins; C557617:GAL4 protein, Drosophila; D034741:RNA, Small Interfering; D014157:Transcription Factors