Retroviral-mediated insertional mutagenesis in zebrafish.

摘要: Abstract Since the initial publication of this chapter in 2004, additional methodologies have been developed which could improve and/or complement original retroviral-mediated insertional mutagenesis. Retroviral vectors also shown to be useful for goals other than In addition, mutagenesis has applied zebrafish use reverse genetics as well forward screening. Finally, mutant collection described herein screened by a number labs find host mutants (with genes already identified) with developmental growth defects affecting eye, liver, skin, craniofacial skeleton, kidney, myeloid cells, hematopoietic stem and axon pathfinding, cell cycle or DNA damage response, altered aging properties, modulated cardiac repolarization. The major complementary approaches new uses technique include: • Pseudotyped retroviruses used deliver enhancer trap vectors, allows selection insertions near particular expression patterns. Although mutagenicity these yet determined, they are purely because generate large transgenic lines visible reporters (e.g., Green Fluorescent Protein GFP) expressed interesting patterns provide information regarding gene regulation context genomic organization. Gain-of-function designed allow dominant genetic screens. Thus, whose overexpression results phenotypes interest can efficiently identified. Retroviruses make library hundreds thousands mapped recovered from frozen sperm samples. Such libraries serve on-the-shelf resources, whereby one obtain mutation nearly any simply recovering an insertion that sperm, similar gene-trap genome-wide targeting ES cells mouse. Transposons effective transgenesis thus may screens – both traps traps. It is possible transposons different site biases, making them important technologies