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1. 发明申请

WO2014100062A1 RECOMBINANT MICROBIAL CELLS THAT PRODUCE AT LEAST 28% EICOSAPENTAENOIC ACID AS DRY CELL WEIGHT 审中-公开
标题翻译：产生至少28％的重要脂肪酸作为干细胞重量的重组微生物细胞
公开(公告)号：WO2014100062A1
公开(公告)日：2014-06-26
申请号：PCT/US2013/075896
申请日：2013-12-18
申请人： E. I. DU PONT DE NEMOURS AND COMPANY
发明人： ZHU, Quinn, Qun , HONG, Seung-Pyo , SHARPE, Pamela, L. , YADAV, Narendra, S. , FAN, Xiaochun , DAUNER, Michael
IPC分类号： C12P7/64 , C12N9/10
CPC分类号： C12P7/6427 , C12N1/16 , C12N9/1029 , C12P7/6463 , C12P7/6472 , C12Y203/01023 , Y02P20/52
摘要： Recombinant microbial cells are disclosed herein that produce an oil comprising at least 28 percent eicosapentaenoic acid (EPA) measured as a weight percent of dry cell weight. These cells may comprise a polynucleotide sequence encoding an active acyl-CoA:lysophosphatidylcholine acyltransferase (LPCAT) comprising at least one amino acid mutation in a membrane-bound O-acyltransferase motif. In addition, the cells may comprise a down-regulation of an endogenous polynucleotide sequence encoding Sou2 sorbitol utilization protein, and/or one or more polynucleotides encoding phospholipid:diacylglycerol acyltransferase (PDAT), delta-12 desaturase, a dihomo-gamma-linolenic acid (DGLA) synthase multizyme, delta-8 desaturase, malonyl-CoA synthetase (MCS), or acyl-CoA:lysophosphatidic acid acyltransferase (LPAAT). Also disclosed are methods of using the recombinant microbial cells to produce oil containing omega-3 polyunsaturated fatty acids such as EPA.
摘要翻译：本文公开了重组微生物细胞，其产生包含以干细胞重量的重量百分比测量的至少28％的二十碳五烯酸（EPA）的油。这些细胞可以包含编码活性酰基-CoA：溶血卵磷脂酰胆碱酰基转移酶（LPCAT）的多核苷酸序列，其包含膜结合的O-酰基转移酶基序中的至少一个氨基酸突变。此外，细胞可以包含编码Sou2山梨糖醇利用蛋白的内源多核苷酸序列和/或编码磷脂：二酰基甘油酰基转移酶（PDAT），δ-12去饱和酶，二氢-γ-亚麻酸的一种或多种多核苷酸的下调（DGLA）合酶多酶，δ-8去饱和酶，丙二酰辅酶A合成酶（MCS）或酰基辅酶A：溶血磷脂酸酰基转移酶（LPAAT）。还公开了使用重组微生物细胞产生含有ω-3多不饱和脂肪酸如EPA的油的方法。

2. 发明申请

WO2016073433A1 PEPTIDE-MEDIATED DELIVERY OF RNA-GUIDED ENDONUCLEASE INTO CELLS 审中-公开
标题翻译：肽介导的RNA引导的内切酶进入细胞
公开(公告)号：WO2016073433A1
公开(公告)日：2016-05-12
申请号：PCT/US2015/058760
申请日：2015-11-03
申请人： E. I. DU PONT DE NEMOURS AND COMPANY
发明人： FRISCH, Ryan L. , FAN, Xiaochun , HONG, Seung-Pyo , JACKSON, Ethel Noland
IPC分类号： C12N9/22 , C12N15/62
CPC分类号： C12N9/22 , C07K2319/09 , C07K2319/10 , C12N9/96 , C12N15/102 , C12N15/62 , C12N15/86 , C12N15/907 , C12N2310/3513 , C12N2750/14141
摘要： A composition is disclosed that comprises at least one protein component of an RNA-guided endonuclease (RGEN) and at least one cell-penetrating peptide (CPP), wherein the RGEN protein component and CPP are covalently or non-covalently linked to each other in an RGEN protein-CPP complex. The RGEN protein-CPP complex can traverse (i) a cell membrane, or (ii) a cell wall and cell membrane, of a cell. The RGEN protein component of an RGEN protein-CPP complex in certain embodiments can be associated with a suitable RNA component to provide an RGEN capable of specific DNA targeting. Further disclosed are compositions comprising at least one protein component of a guide polynucleotide/Cas endonuclease complex and at least one CPP, as well as methods of delivering RGEN proteins into microbial cells, as well as methods of targeting DNA with RGENs.
摘要翻译：公开了包含RNA引导的内切核酸酶（RGEN）和至少一种细胞穿透肽（CPP）的至少一种蛋白质组分的组合物，其中所述RGEN蛋白质组分和CPP彼此共价或非共价连接 RGEN蛋白-CPP复合物。 RGEN蛋白-CPP复合物可以穿过（i）细胞膜，或（ii）细胞的细胞壁和细胞膜。在一些实施方案中，RGEN蛋白-CPP复合物的RGEN蛋白组分可以与合适的RNA组分相关联，以提供能够进行特异性DNA靶向的RGEN。还公开了包含导向多核苷酸/ Cas内切核酸酶复合物和至少一种CPP的至少一种蛋白质组分的组合物，以及将RGEN蛋白递送到微生物细胞中的方法，以及用RGEN靶向DNA的方法。

3. 发明申请

WO2016025131A1 GENETIC TARGETING IN NON-CONVENTIONAL YEAST USING AN RNA-GUIDED ENDONUCLEASE 审中-公开
标题翻译：使用RNA指导的内切酶在非常规的YEAST中进行遗传定位
公开(公告)号：WO2016025131A1
公开(公告)日：2016-02-18
申请号：PCT/US2015/041256
申请日：2015-07-21
申请人： E. I. DU PONT DE NEMOURS AND COMPANY
发明人： FRISCH, Ryan , FAN, Xiaochun , HONG, Seung-Pyo
IPC分类号： C12N9/16 , C12N15/10 , C12N15/113
CPC分类号： C12N15/905 , C12N9/16 , C12N15/102 , C12N15/113 , C12N2310/121 , C12N2310/20
摘要： Non-conventional yeasts are disclosed herein comprising at least one RNA-guided endonuclease (RGEN) comprising at least one RNA component that does not have a 5'-cap. This uncapped RNA component comprises a sequence complementary to a target site sequence in a chromosome or episome in the yeast. The RGEN can bind to, and optionally cleave, one or both DNA strands at the target site sequence. An example of an RGEN herein is a complex of a Cas9 protein with a guide RNA. A ribozyme is used in certain embodiments to provide an RNA component lacking a 5'-cap. Further disclosed are methods of genetic targeting in non-conventional yeast.
摘要翻译：本文公开了包含至少一种RNA引导内切核酸酶（RGEN）的非常规酵母，其包含至少一种不具有5'帽的RNA成分。该未加帽的RNA组分包含与酵母中的染色体或附加体中的靶位点序列互补的序列。 RGEN可以在目标位点序列上结合并任选地切割一条或两条DNA链。本文的RGEN的实例是Cas9蛋白与引导RNA的复合物。在某些实施方案中使用核酶提供缺少5'-帽的RNA成分。进一步公开的是在非常规酵母中遗传靶向的方法。

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