Culture of drosophila primary cells dissociated from gastrula embryos and their use in rnai screening

Nature Protocols

Volumn 4, Issue 10, 2009, Pages 1502-1512

  Bai, Jianwu ^a   Sepp, Katharine J ^a   Perrimon, Norbert ^a,b  

^a Department of Genetics ^*  (United States)

^b HOWARD HUGHES MEDICAL INSTITUTE   (United States)

Author keywords

[No Author keywords available]

Indexed keywords

DOUBLE STRANDED RNA;

ANIMAL; ARTICLE; CELL CULTURE; CULTURE MEDIUM; CULTURE TECHNIQUE; CYTOLOGY; DROSOPHILA; GASTRULA; GENOMICS; INSTRUMENTATION; PRENATAL DEVELOPMENT; RNA INTERFERENCE;

ANIMALS; CELL CULTURE TECHNIQUES; CELLS, CULTURED; CULTURE MEDIA; DROSOPHILA; GASTRULA; GENOMICS; RNA INTERFERENCE; RNA, DOUBLE-STRANDED;

EID: 73349091855     PISSN: 17542189     EISSN: 17502799     Source Type: Journal    
DOI: 10.1038/nprot.2009.147     Document Type: Article

References (24)

1. Adams, M.D. et al. The genome sequence of Drosophila melanogaster. Science 287, 2185-2195 (2000).
2. Perrimon, N. & Mathey-Prevota, B. Applications of high-throughput RNA interference screens to problems in celland developmental biology. Genetics 175, 7-16 (2007).
3. Steinbrink, S. & Boutros, M. RNAi screening in cultured Drosophila cells. Methods Mol. Biol. 420, 139-153 (2008).
4. Bai, J., Hartwig, J.H. & Perrimon, N. SALS, a WH2-domain-containing protein, promotes sarcomeric actin filament elongation from pointed ends during Drosophila muscle growth. Dev. Cell 13, 828-842 (2007).
5. Bai, J. et al. RNA interference screening in Drosophila primary cells for genes involved in muscle assembly and maintenance. Development 135, 1439-1449 (2008).
6. Sepp, K.J. et al. Identification of neural outgrowth genes using genome-wide RNAi. PloS. Genet. 4, e1000111 (2008).
7. Echalier, G. in Drosophila Cells in Culture (ed. Echalier, G.) 71-127 (Academic Press, London, 1997).
8. Donady, J.J. & Fyrberg, E. Mass culturing of Drosophila embryonic cells in vitro. TCA Manual 3, 685-687 (1977).
9. Seecof, R.L., Alleaume, N., Teplitz, R.L. & Gerson, I. Differentiation of neurons and myocytes in cell cultures made from Drosophila gastrulae. Exp. Cell Res. 69, 161-173 (1971).
10. Kuppers-Munther, B. et al. A new culturing strategy optimises Drosophila primary cell cultures for structural and functional analyses. Dev. Biol. 269, 459-478 (2004).
11. Donady, J. & Seecof, R. Effect of the gene lethal (1) myospheroid on Drosophila embryonic cells in vitro. In Vitro 8, 7-12 (1972).
12. Noll, E., Perkins, L., Mahowald, A. & Perrimon, N. Approaches to identify genes involved in Drosophila embryonic CNS development. J. Neurobiol. 24, 701-722 (1993).
13. Storti, R., Horovitch, S., Scott, M., Rich, A. & Pardue, M. Myogenesis in primary cell cultures from Drosophila melanogaster. protein synthesis and actin heterogeneity during development. Cell 13, 589-598 (1978).
14. Simcox, A. et al. Efficient genetic method for establishing Drosophila cells lines unlocks the potential to create lines of specific genotypes. PLoS. Genet. 4, e1000142 (2008).
15. Estrada, B. et al. An integrated strategy for analyzing the unique developmental programs of different myoblast subtypes. PloS. Genet. 2, e16 (2006).
16. Brand, A. & Perrimon, N. Targeted gene expression as a means of altering cell fates and generating dominant phenotypes. Development 118, 401-415 (1993).
17. Shields, G. & Sang, J. Characteristics of five cell types appearing during in vitro culture of embryonic material from Drosophila melanogaster. J. Embryol. Exp. Morph. 23, 53-69 (1970).
18. Shields, G., Dubendorfer, A. & Sang, J. Differentiation in vitro of larval cell types from early embryonic cells of Drosophila melanogaster. J. Embryol. Exp. Morphol. 33, 159-175 (1975).
19. Hayashi, I. & Perez-Magallanes, M. Establishment of pure neuronal and muscle precursor cell cultures from Drosophila early gastrula stage embryos. In Vitro Cell Dev. Biol. Anim. 30A, 202-208 (1994).
20. Fogerty, F.J. et al. Tiggrin, a novel Drosophila extracellular matrix protein that functions as a ligand for Drosophila PS2PS integrins. Development 120, 1747-1758 (1994).
21. Ramadan, N., Flockhart, I., Brooker, M., Perrimon, N. & Mathey-Prevot, B. Design and implementation of high-throughput RNAi screensin cultured Drosophila cells. Nat. Protoc. 2, 2245-2264 (2007).
22. Clemens, J. et al. Use ofdouble-stranded RNA interference in Drosophila cell lines to dissectsignaltransduction pathways. Proc. Natl. Acad. Sci. USA 97, 6499-6503 (2000).
23. Macleod, G., Suster, M., Charlton, M. & Artwood, H. Single neuron activity in the Drosophila larval CNS detected with calcium indicators. J. Neurosci. Methods 127, 167-178 (2003).
24. Ashburner, M. & Roote, J. Laboratory culture of Drosophila. (eds. Sullivan, W., Ashburner, M. & Hawley, R.S.) 588-599. (Cold Spring Harbor Laboratory Press, Cold Spring Harbor, New York, 1999).