Gene Stacking in Plants Through the Application of Site-Specific Recombination and Nuclease Activity.
Gene Stacking in Plants Through the Application of Site-Specific Recombination and Nuclease Activity.
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通过应用位点特异性重组和核酸酶活性进行植物基因堆积。
DOI：
10.1007/978-1-4939-8778-8_18
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发表时间：
2018
影响因子：
--
通讯作者：
V. Srivastava
中科院分区：
文献类型：
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作者：
V. Srivastava
为了将有价值的性状引入作物品种，需要将基因一个接一个或作为一个片段插入植物基因组的生物技术方法。将多个基因插入到一个位点，称为分子堆叠，对于允许基因共同遗传到后代是很重要的。一般来说，有两种方法可用于创建基因堆栈：核酸酶诱导的靶向基因整合到天然位点和重组酶介导的基因整合到工程位点。由于重组酶在植物基因组中具有较高的效率和精度，其应用前景十分广阔。本章描述了一种基于cre -loxsite特异性重组系统的基因堆叠方法，该方法将基因整合到工程位点和核酸酶中，以删除选择基因，导致性状堆叠到单个基因组位点。Cre-loxin在许多植物物种中的高效、精确和不可检测的脱靶效应，使其成为基因堆叠等复杂应用的有吸引力的工具。
Biotechnology methods for inserting genes one by one or as a block of fragment into plant genomes are needed to introduce valuable traits into crop varieties. Insertion of multiple genes into a single site, called as molecular stacking, is important to allow co-inheritance of the genes into the progeny. Generally, two approaches are available for creating gene stacks: nuclease-induced targeted gene integration into native sites and recombinase-mediated gene integration into the engineered sites. The recombinase application is attractive as several recombinases show high efficiency and precision in plant genomes. This chapter describes a gene stacking method based on the use of Cre-loxsite-specific recombination system to integrate genes into the engineered sites and nucleases to delete selection genes leading to stacking of traits into a single genomic site. High efficiency and precision, and undetectable off-target effects of Cre-loxin a number of plant species, make it an attractive tool for complex applications such as gene stacking.