Forward Genetic Screen Using Zebrafish to Identify New Genes Involved in Myelination.

Forward Genetic Screen Using Zebrafish to Identify New Genes Involved in Myelination.
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使用斑马鱼进行正向遗传筛选来识别参与髓鞘形成的新基因。

DOI:
10.1007/978-1-4939-9072-6_11
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发表时间:
2019
期刊:
Methods in molecular biology (Clifton, N.J.)
影响因子:
--
通讯作者:
Kegel L
Kegel L
中科院分区:
--
文献类型:
--
作者:
Kegel L

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斑马鱼现在被公认为是卓越的脊椎动物模型,用来进行基因发现/正向遗传筛选,以确定生物过程的分子遗传学基础。斑马鱼的基因发现筛选已经为神经胶质细胞的发育和功能机制提供了新的见解。斑马鱼的绝大多数遗传筛查都是基于三代筛查,首先使用化学诱变剂ENU随机诱导成年雄性斑马鱼的突变。在这里,我们概述了支持这类筛选的方法,详细说明了从ENU突变到在后续世代中恢复纯合突变动物所需的育种方案、筛选程序本身,重点是髓鞘胶质细胞的分析,以及随后突变表型的确认。
Zebrafish are now well established as the preeminent vertebrate model with which to carry out gene discovery/forward genetic screens to identify the molecular genetic basis of biological processes. Gene discovery screens in zebrafish have already provided novel insight into mechanisms of glial cell development and function. The vast majority of genetic screens in zebrafish are based around a three generation screen that starts with the random induction of mutations in adult males using the chemical mutagen ENU. Here we outline the methods that underlie this type of screen, detailing each step, from ENU mutagenesis, through the breeding schemes required to recover homozygous mutant animals in subsequent generations, the screening procedure itself, with a focus on the analysis of myelinating glia, and the subsequent confirmation of mutant phenotypes.
DOI: 10.1038/nbt.2501
发表时间: 2013-03
影响因子: 46.9
作者:
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发表时间: 1997
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