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    • 7. 发明申请
    • FLUORESCENCE ACTIVATED CELL SORTING (FACS) ENRICHMENT TO GENERATE PLANTS
    • 荧光激活细胞分选(FACS)生产植物
    • WO2014039970A1
    • 2014-03-13
    • PCT/US2013/058766
    • 2013-09-09
    • DOW AGROSCIENCES LLC
    • SPANGENBERG, GermanSAHAB, SareenaMASON, John
    • C12N15/82C12Q1/24A01H5/00
    • C12N15/8241C12N15/8212C12N15/8213
    • An Engineered Transgene Integration Platform (ETIP) is described that can be inserted randomly or at targeted locations in plant genomes to facilitate rapid selection and detection of a GOI that is perfectly targeted (both the 3? and 5? ends) at the ETIP genomic location. One element in the invention is the introduction of specific double stranded breaks within the ETIP. In some embodiments, an ETIP is described using zinc finger nuclease binding sites, but may utilize other targeting technologies such as meganucleases, TALs, CRISPRs, or leucine zippers. Also described are compositions of, and methods for producing, transgenic plants wherein the donor or payload DNA expresses one or more products of an exogenous nucleic acid sequence (e.g. protein or RNA) that has been stably-integrated into an ETIP in a plant cell. In embodiments, the ETIP facilitates testing of gene candidates and plant expression vectors from ideation through Development phases.
    • 描述了可以随机插入植物基因组中的目标位置的工程化转基因整合平台(ETIP),以便于在ETIP基因组位置上完全靶向(3'和5'末端)的GOI的快速选择和检测 。 本发明中的一个要素是在ETIP内引入特定的双链断裂。 在一些实施方案中,使用锌指核酸酶结合位点描述ETIP,但可以利用其它靶向技术,例如大范围核酸酶,TAL,CRISPR或亮氨酸拉链。 还描述了转基因植物的组合物和生产方法,其中供体或有效载体DNA表达已经稳定整合到植物细胞中的ETIP中的外源核酸序列(例如蛋白质或RNA)的一种或多种产物。 在实施方案中,ETIP有助于从构想到开发阶段测试基因候选物和植物表达载体。