公开(公告)号：US20220325303A1

公开(公告)日：2022-10-13

申请号：US17840369

申请日：2022-06-14

Applicant: AMYRIS, INC.

Inventor： Andrew HORWITZ ,  Kristy Michelle HAWKINS ,  Max SCHUBERT ,  Wayne SZETO

IPC: C12N15/90 ,  C12Q1/6897 ,  C12N15/11

Abstract: Provided herein are methods of integrating one or more exogenous nucleic acids into one or more selected target sites of a host cell genome. In certain embodiments, the methods comprise contacting the host cell genome with one or more integration polynucleotides comprising an exogenous nucleic acid to be integrated into a genomic target site, a nuclease capable of causing a break at the genomic target site, and a linear nucleic acid capable of homologous recombination with itself or with one or more additional linear nucleic acids contacted with the population of cells, whereupon said homologous recombination results in formation of a circular extrachromosomal nucleic acid comprising a coding sequence for a selectable marker. In some embodiments, the methods further comprise selecting a host cell that expresses the selectable marker.

公开(公告)号：US20150093797A1

公开(公告)日：2015-04-02

申请号：US14474976

申请日：2014-09-02

Applicant: AMYRIS, INC.

Inventor： Timothy Stevens GARDNER ,  Kristy Michelle HAWKINS ,  Adam Leon MEADOWS ,  Annie Ening TSONG ,  Yoseph TSEGAYE

IPC: C12P5/00 ,  C12N15/81

CPC classification number: C12P5/007 ,  C12N9/0006 ,  C12N9/0008 ,  C12N9/1288 ,  C12N15/52 ,  C12N15/81 ,  C12P7/42 ,  C12P23/00 ,  C12Y101/01034 ,  C12Y102/0101 ,  C12Y207/08

Abstract: Provided herein are compositions and methods for the heterologous production of acetyl-CoA-derived isoprenoids in a host cell. In some embodiments, the host cell is genetically modified to comprise a heterologous nucleotide sequence encoding an acetaldehyde dehydrogenase, acetylating (ADA, E.C. 1.2.1.10) and an MEV pathway comprising an NADH-using HMG-CoA reductase. In some embodiments, the host cell is genetically modified to comprise a heterologous nucleotide sequence encoding an ADA and an MEV pathway comprising an acetoacetyl-CoA synthase. In some embodiments, the genetically modified host cell further comprises one or more heterologous nucleotide sequences encoding a phosphoketolase and a phosphotransacetylase. In some embodiments, the genetically modified host cell further comprises a functional disruption of the native PDH-bypass. The compositions and methods described herein provide an energy-efficient yet redox balanced route for the heterologous production of acetyl-CoA-derived isoprenoids.

Abstract translation: 本文提供了在宿主细胞中异源产生乙酰辅酶A衍生的类异戊二烯的组合物和方法。 在一些实施方案中，宿主细胞被遗传修饰以包含编码乙醛脱氢酶，乙酰化（ADA，E.C.1.2.1.10）和包含使用NADH的HMG-CoA还原酶的MEV途径的异源核苷酸序列。 在一些实施方案中，宿主细胞被遗传修饰以包含编码包含乙酰乙酰辅酶A合成酶的ADA和MEV途径的异源核苷酸序列。 在一些实施方案中，经遗传修饰的宿主细胞还包含一个或多个编码磷酸酮醇酶和磷酸转乙酰酶的异源核苷酸序列。 在一些实施方案中，遗传修饰的宿主细胞还包括天然PDH旁路的功能性破坏。 本文描述的组合物和方法提供了用于异源产生乙酰辅酶A衍生的类异戊二烯的能量效率尚未的氧化还原平衡途径。

公开(公告)号：US20130236942A1

公开(公告)日：2013-09-12

申请号：US13752293

申请日：2013-01-28

Applicant: AMYRIS, INC.

Inventor： Timothy Stevens GARDNER ,  Kristy Michelle HAWKINS ,  Adam Leon MEADOWS ,  Annie Ening TSONG ,  Yoseph TSEGAYE

IPC: C12P5/00 ,  C12N15/81

CPC classification number: C12P5/007 ,  C12N9/0006 ,  C12N9/0008 ,  C12N9/1288 ,  C12N15/52 ,  C12N15/81 ,  C12P7/42 ,  C12P23/00 ,  C12Y101/01034 ,  C12Y102/0101 ,  C12Y207/08

Abstract: Provided herein are compositions and methods for the heterologous production of acetyl-CoA-derived isoprenoids in a host cell. In some embodiments, the host cell is genetically modified to comprise a heterologous nucleotide sequence encoding an acetaldehyde dehydrogenase, acetylating (ADA, E.C. 1.2.1.10) and an MEV pathway comprising an NADH-using HMG-CoA reductase. In some embodiments, the host cell is genetically modified to comprise a heterologous nucleotide sequence encoding an ADA and an MEV pathway comprising an acetoacetyl-CoA synthase. In some embodiments, the genetically modified host cell further comprises one or more heterologous nucleotide sequences encoding a phosphoketolase and a phosphotransacetylase. In some embodiments, the genetically modified host cell further comprises a functional disruption of the native PDH-bypass. The compositions and methods described herein provide an energy-efficient yet redox balanced route for the heterologous production of acetyl-CoA-derived isoprenoids.