Unsuccessful attempt at gene-editing by homologous recombination in the zebrafish germ line using the approach of "Rong and Golic".

Unsuccessful attempt at gene-editing by homologous recombination in the zebrafish germ line using the approach of "Rong and Golic".
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DOI:
10.1007/s11248-012-9607-1
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发表时间:
2012-10
影响因子:
3
通讯作者:
Brown W
Brown W
中科院分区:
生物学4区
文献类型:
--
作者:
Brookfield R;Dafhnis-Calas F;Xu Z;Brown W

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我们已经研究了在斑马鱼中实施通过同源重组进行位点特异性编辑的策略的实用性,该策略类似于Rong和Golic(Rong和Golic in Genetics 157:1307-1312)在果蝇中开发的策略。我们分析了来自22个杂交的大约7,300个后代,并证明了基因编辑构建体的成功切除,但未能检测到基因编辑或切除后完整编辑构建体的随机整合。在我们的数据集中的事件的聚类表明,切除事件不是独立发生的,并强调,启动子驱动高水平,在减数分裂细胞中的组织特异性转录可能是必要的,如果通过同源重组的位点特异性编辑的一般方法是履行其潜力。本文的在线版本(doi:10.1007/s11248-012-9607-1)包含补充材料,可供授权用户使用。
We have investigated the practicality of implementing a strategy for site-specific editing by homologous recombination in zebrafish analogous to that developed by Rong and Golic (Rong and Golic in Genetics 157:1307–1312,) in Drosophila melanogaster. We analysed approximately 7,300 offspring from 22 crosses and demonstrated successful excision of the gene editing construct but failed to detect either gene editing or the random integration of the intact editing construct subsequent to excision. The clustering of events in our data set demonstrates that the excision events are not occurring independently and emphasise that a promoter driving high level, tissue-specific transcription in meiotic cells is likely to be necessary if this general approach to site-specific editing by homologous recombination is to fulfil its potential. The online version of this article (doi:10.1007/s11248-012-9607-1) contains supplementary material, which is available to authorized users.
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