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Chapters
- 00:05Title
- 01:20Bioinformatics and Expressions Analysis in the Developing Zebrafish Embryo
- 02:27Site-directed Mutagenesis and in vitro mRNA Transcription
- 03:54In vivo Assay of Variant Loss of Function
- 06:40In vivo Assay of Variant Dominant Negative or Gain of Function Effects
- 08:11Representative MO Knockdown and Rescue Results
- 12:06Conclusion
Here, we present a systematic approach for developing physiologically relevant, sensitive and specific in vivo assays for interpreting variation in human pathology. Transient genetic manipulation via microinjection of WT and mutant human mRNA and morpholino (MO) antisense oligonucleotides harness the tractability of the developing zebrafish embryo to rapidly assay pathogenic mutations, especially, but not exclusively, in the context of human developmental disorders.
