Once mutations are identified, the mutant zebrafish line is recovered through in vitro fertilization of wild-type eggs using the corresponding cryopreserved sperm samples. Mutational screening can be performed either by direct sequencing of the gene of interest or through the use of the CEL I nuclease, an enzyme that recognizes and cuts DNA heteroduplexes to identify single base pair differences between the wild-type and mutant alleles of the target gene. Sperm from F1 males is cryopreserved for future reconstitution of the mutant line once relevant mutations are identified, while genomic DNA from these animals is isolated for screening. ENU mutagenesis is performed on male fish that are bred to wild-type females. Also known as TILLING (Targeting Induced Local Lesions In Genomes), this process utilizes DNA from mutagenized fish to screen directly for mutations of zebrafish genes of interest. Background and techniquesĪnother recently developed technique employs target-selected mutagenesis in zebrafish. This review will highlight some of the advantages of the zebrafish system, introduce the technologies that can be used to study myeloid cell development in the zebrafish, and highlight some of the recent observations and discoveries that have contributed to our understanding of myeloid cell biology. Due to its inherent advantages as a developmental model, including external fertilization and optical clarity, and through the use of specific technologies, such as RNA in situ hybridization, morpholino-induced gene knockdown, and mutant and transgenic fish lines, the zebrafish has helped provide important insights into the genetic regulation of vertebrate myelopoiesis. Zebrafish hematopoiesis has been shown to be very similar to that of higher vertebrates, and homologues of a large number of genes involved in mammalian myelopoiesis have been identified in the zebrafish. The tropical freshwater fish, Danio rerio, provides a powerful model organism for studying the molecular control of normal and abnormal myelopoiesis.
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