利用通用盒式穿梭选择策略构建小鼠全长平衡染色体

Generation of a Mouse Full-length Balancer with Versatile Cassette-shuttling Selection Strategy.

作者信息

Ye Zhisheng, Sun Lei, Li Rongbo, Han Min, Zhuang Yuan, Wu Xiaohui, Xu Tian

机构信息

1. State Key Laboratory of Genetic Engineering and National Center for International Research of Development and Disease, Collaborative Innovation Center for Genetics and Development, Fudan-Yale Biomedical Research Center, Institute of Developmental Biology and Molecular Medicine, Fudan University, Shanghai 200433, China.; 2. Howard Hughes Medical Institute, Department of Molecular, Cellular, Developmental Biology, University of Colorado at Boulder, Boulder, CO 80309, USA.

出版信息

Int J Biol Sci. 2016 Jun 7;12(8):911-6. doi: 10.7150/ijbs.15172. eCollection 2016.

DOI:10.7150/ijbs.15172

PMID:27489495

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC4971730/

Abstract

Balancer chromosomes are important tools for a variety of genetic manipulations in lower model organisms, owing to their ability to suppress recombination. In mouse, however, such effort has not been accomplished, mostly due to the size of the chromosomes and the complexity of multiple step chromosomal engineering. We developed an effective and versatile cassette-shuttling selection (CASS) strategy involving only two selection markers to achieve the sequential production of multiple large inversions along the chromosome. Using this strategy, we successfully generated the first full-length balancer in mice and showed that Balancer 17M-GFP can efficiently suppress recombination. Our study has not only generated a useful genetic resource, but also provided a strategy for constructing mammalian balancer chromosomes.

摘要

平衡染色体是在低等模式生物中进行各种基因操作的重要工具，因为它们能够抑制重组。然而，在小鼠中，这项工作尚未完成，主要是由于染色体的大小以及多步染色体工程的复杂性。我们开发了一种有效且通用的盒式穿梭选择（CASS）策略，该策略仅涉及两个选择标记，以实现沿染色体顺序产生多个大的倒位。使用这种策略，我们成功地在小鼠中产生了第一个全长平衡染色体，并表明平衡染色体17M-GFP能够有效地抑制重组。我们的研究不仅产生了一种有用的遗传资源，还提供了一种构建哺乳动物平衡染色体的策略。