Multiplex Detection of RNA Expression in Drosophila Embryos

@article{Kosman2004MultiplexDO,
  title={Multiplex Detection of RNA Expression in Drosophila Embryos},
  author={David Kosman and Claudia Mieko Mizutani and Derek Lemons and W Gregory Cox and William McGinnis and Ethan Bier},
  journal={Science},
  year={2004},
  volume={305},
  pages={846 - 846}
}
We present a fluorescence-based, multiplex in situ hybridization method that permits the simultaneous detection of five differently labeled antisense RNA probes and up to seven different transcripts in a single Drosophila embryo. We also show that it should be possible to increase the number of detected transcripts substantially with nascent transcript multiplex fluorescent in situ hybridization. These multiplex methods fill a current technological gap between high-resolution in situ… Expand
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References

SHOWING 1-10 OF 15 REFERENCES
A non-radioactive in situ hybridization method for the localization of specific RNAs in Drosophila embryos reveals translational control of the segmentation gene hunchback
TLDR
A non-radioactive in situ hybridization technique for the localization of RNA in whole mount Drosophila embryos and revealed translational control of the maternally derived hb mRNA, which was difficult to detect by conventional techniques. Expand
Single-Cell Gene Expression Profiling
TLDR
By combining advances in computational fluorescence microscopy with multiplex probe design, this work devised technology in which the expression of many genes can be visualized simultaneously inside single cells with high spatial and temporal resolution. Expand
Systematic determination of patterns of gene expression during Drosophila embryogenesis
TLDR
Analyzing gene-expression patterns by in situ hybridization to whole-mount embryos provides an extremely rich dataset that can be used to identify genes involved in developmental processes that have been missed by traditional genetic analysis. Expand
Whole-Genome Analysis of Dorsal-Ventral Patterning in the Drosophila Embryo
TLDR
Whole-genome microarray assays identified as many as 40 new Dorsal target genes, which encode a broad spectrum of cell signaling proteins and transcription factors, which represent one of the most extensive gene networks known for any developmental process. Expand
Transcribed genes are localized according to chromosomal position within polarized Drosophila embryonic nuclei
TLDR
Using a high-resolution in situ hybridization method, it is found that each of 15 transcribed genes was localized as predicted by their chromosomal position and by the known polarized organization of the chromosomes. Expand
Directing cell division during development.
TLDR
After this transition to zygotic control, pulses of string transcription define the timing of highly patterned embryonic cell divisions and cyclin accumulation is not rate limiting. Expand
Dynamic control of positional information in the early Drosophila embryo
TLDR
This analysis implies that the threshold-dependent interpretation of maternal morphogen concentration is not sufficient to determine shifting gap domain boundary positions, and suggests that establishing and interpreting positional information are not independent processes in the Drosophila blastoderm. Expand
Supporting Online Material www.sciencemag.org/cgi/content/full
  • DC1 Materials and Methods SOM Text
  • 2004
Genome Biol
  • Genome Biol
  • 2002
Curr. Biol
  • Curr. Biol
  • 1999
...
1
2
...