A Transcriptome-wide RNAi Screen in the Drosophila Ovary Reveals Factors of the Germline piRNA Pathway
Open accessAbstract
The Drosophila piRNA pathway provides an RNA-based immune system that defends the germline genome against selfish genetic elements. Two interrelated branches of the piRNA system exist: somatic cells that support oogenesis only employ Piwi, whereas germ cells utilize a more elaborate pathway centered on the three gonad-specific Argonaute proteins (Piwi, Aubergine, and Argonaute 3). While several key factors of each branch have been identified, our current knowledge is insufficient to explain the complex workings of the piRNA machinery. Here, we report a reverse genetic screen spanning the ovarian transcriptome in an attempt to uncover the full repertoire of genes required for piRNA-mediated transposon silencing in the female germline. Our screen reveals key factors of piRNA-mediated transposon silencing, including the piRNA biogenesis factors CG2183 (GASZ) and Deadlock. Our data uncover a previously unanticipated set of factors preferentially required for repression of different transposon types.
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For the remaining 260 genes, the stocks used an alternative integration site that is not problematic.
“For the remaining 260 genes, the stocks used the alternative integration site which is not problematic, with KK stocks having been used successfully in a range of different screens [29,34].”
From Functional unknomics: Systematic screening of conserved genes of unknown function (2023) · cites this paper as [34]
The Results section describes screening VDRC RNAi lines, and Experimental Procedures refers stock details to the Extended Experimental Procedures. Searches for “260,” “KK,” and “integration site” found no passage in our copy establishing that the remaining 260 genes used a nonproblematic alternative site. (LLM, conf=0.74)
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Self-citation
- Authors
- Czech, B., Preall, J. B., McGinn, J., & Hannon, G. J.
- Affiliations
- Howard Hughes Medical Institute (US)
- Corresponding author
- Czech, Benjamin
- Hannon, Gregory J.
- Venue
- Molecular Cell
- Publisher
- Elsevier BV
- Published
- 2013-06
- Volume
- 50
- Issue
- 5
- Pages
- 749-761
- ISSN
- 1097-2765 (ISSN-L)
- PMC
- PMC3724427
- OpenAlex
- W1997570561
- Cited by
- 269
- References
- 62
- Citations by year
- 2026: 8 · 2025: 24 · 2024: 16 · 2023: 20 · 2022: 11 · 2021: 26 · 2020: 23 · 2019: 21 · 2018: 24 · 2017: 21 · 2016: 16 · 2015: 25 · 2014: 26 · 2013: 8
- FWCI
- 30.19
- Type
- journal article
- Language
- en
- Open access
- bronze
- doi.org
- Licence
- Locations
- Molecular Cell (publishedVersion)
- PubMed (publishedVersion)
- Europe PMC (PubMed Central) (acceptedVersion)
- PubMed Central (acceptedVersion)
- Cold Spring Harbor Laboratory Institutional Repository (Cold Spring Harbor Laboratory) (acceptedVersion)
- Funders
- Howard Hughes Medical Institute
- American Cancer Society
- Boehringer Ingelheim Fonds
- National Institutes of Health
- National Institute of General Medical Sciences (R01GM062534, R37GM062534)
- Topics
- Life Sciences › Agricultural and Biological Sciences › Plant Science › Chromosomal and Genetic Variations
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › RNA Interference and Gene Delivery
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Epigenetics and DNA Methylation
- Keywords
- piRNA pathway, germline, transposon silencing, RNAi screen, Drosophila ovary, Piwi, eggplant, deadlock, selfish genetic elements
- MeSH terms
- Animals, DNA Transposable Elements, Drosophila melanogaster, Female, Microtubule-Associated Proteins, Ovary, Peptide Initiation Factors, Reproducibility of Results, Gene Silencing, Gene Expression Profiling, Drosophila Proteins, RNA Interference, RNA, Small Interfering, Argonaute Proteins
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