专利号:US-2025049961-A1 优先权日:2021-12-23 标题 :Scalable and high-purity cell-free synthesis of closed-ended dna vectors 发明人:MONDS RUSSELL; CIPI JORIS; BLACKSTOCK DANIEL JASON; DURANT JOHN CHESTER 权利人:GENERATION BIO CO 摘要:This disclosure provides methods for scalable and high-purity cell-free synthesis of DNA vectors, particularly closed-ended DNA vectors (e.g., ceDNA vectors) having linear and continuous structure for delivery and expression of a transgene. The cell-free synthesis includes digesting a double-stranded DNA construct with at least one restriction endonuclease that is capable of cleaving the construct at cleavage sites, which are distinct from the recognition sites, to release an insert having unique overhangs that regulate the high specificity of the subsequent ligation reaction. The insert is then ligated with inverted terminal repeat (ITR) oligonucleotides to form the closed-ended DNA vector. Corresponding DNA vectors prepared by these methods and related products as well other base and intermediate vectors and constructs associated with the methods are also provided in this disclosure.
专利号:US-12442015-B2 优先权日:2018-01-19 标 题:Closed-ended DNA vectors obtainable from cell-free synthesis and process for obtaining ceDNA vectors 发明人:ALKAN OZAN; KOTIN ROBERT MICHAEL; STANTON MATTHEW; KERR DOUGLAS ANTHONY; PELLETIER CAROLYN 权利人:GENERATION BIO CO 摘要:The application describes methods for synthetic synthesis and cell-free synthesis of DNA vectors, particularly closed-ended DNA vectors (e.g., ceDNA vectors) having linear and continuous structure for delivery and expression of a transgene. The present invention relates to an in vitro process for production of closed-ended DNA vectors, corresponding DNA vector products produced by the methods and uses thereof, and oligonucleotides and kits useful in the process of the invention. DNA vectors produced using the methods described herein are free from unwanted side effects due to contaminants introduced during production in cell lines, for example, bacterial or insect cell lines. Further provided herein are methods and cell lines for reliable gene expression in vitro, ex vivo and in vivo using the ceDNA vectors synthesized using the methods herein.
专利号:US-5455157-A 优先权日:1989-05-22 标 题:Method for the non-radioactive measurement of the nucleic acid synthesis in eukaryotic cells 发明人:HINZPETER MATTHIAS; ELTZ HERBERT VON DER 权利人:BOEHRINGER MANNHEIM GMBH 摘要:For the determination of the rate of nucleic acid synthesis in eukaryotic cells a nucleotide which is non-radioactively labelled or derivatized with a hapten is introduced into the cells with the addition of liposomes and the rate of synthesis of the nucleic acids is determined by means of the incorporation of the nucleotide via its label or derivatization.
专利号:EP-1548006-B1 优先权日:2003-12-12 标题 :Synthesis of tetraazapentamethine derivatives, keratin fibers dyeing process which uses such synthesis 发明人:PEREIRA RUI; BURGAUD HERVE 权利人:OREAL 摘要:Producing tetraazapentamethine compounds (I) comprising reacting an azine compound with an oxidizing agent, is new. Producing tetraazapentamethine compounds of formula (I) comprises reacting an azine compound of formula (VI) or (VII) with an oxidizing agent; [Image] [Image] W1a cationic heteroaromatic group of formula (II) or (III); W2a heteroaromatic group of formula (IV) or (V); Z14Z7 or Z1; Z15Z8 or Z2; Z16Z13 or Z0; R25R5 or R1; Z17Z9 or Z3; Z18Z10 or Z4; Z19Z11 or Z5; Z20Z12 or Z6; R26R8 or R3; R27R7 or R4; R28, R29H, 1-6C alkyl or 1-6C alkoxy, but not both H; Z0CR2, N or NR21; Z1O, S or NR9; Z2N or CR10; Z3N or CR11; Z4N or CR12; Z5N or CR13; Z5N or CR13; Z6N or CR14; Z7O, S or NR15; Z8N or CR16; Z9N or CR17; Z10N or CR18; Z11N or CR19; Z12N or CR20; Z13CR6, N or NR22; R2, R6, R10, R16H; 1-4C alkyl substituted with 0-3 of OH, 1-2C alkoxy, 2-4C (poly)hydroxyalkoxy, NH2, 1-2C (di)alkylamino, COOH or SO3H; phenyl substituted with 0-2 of OH, 1-2C alkoxy, 2-4C (poly)hydroxyalkoxy, NH2, 1-2C (di)alkylamino; COOH; sulfonylamino; R1, R4, R5, R7, R9, R15, R21, R221-6C alkyl substituted with 0-3 of OH, 1-2C alkoxy, 2-4C (poly)hydroxyalkoxy, NH2, 1-2C (di)alkylamino, COOH or SO3H; R0, R3, R8, R11-R14, R17-R20H; 1-6C linear or branched hydrocarbyl with 0-3 unsaturations, optionally substituted with 1-3 of OH, 1-2C alkoxy, 2-4C (poly)hydroxyalkoxy, NH2, 1-2C (di)alkylamino, COOH, SO3H, sulfonylamino, 2-4C (poly)hydroxyalkylamino or halo and optionally interrupted by O, N, S or SO2; phenyl optionally substituted with 1-3 of OH, 1-2C alkoxy, 2-4C (poly)hydroxyalkoxy, NH2, 1-2C (di)alkylamino, COOH, SO3H, sulfonylamino, 2-4C (poly)hydroxyalkylamino or halo; pyrazolyl, pyrrolyl, imidazolyl, thiazolyl, oxazolyl, triazolyl, pyridyl, pyrimidinyl, triazinyl, pyrazinyl or pyridazinyl; X : an anion; R2+R10, R11+R12, R6+R16, R17+R18 can form a 5-6C aromatic ring substituted with 0-2 of OH, NH2, 1-2C (di)alkylamino, 1-2C alkoxy, 2-4C (poly)hydroxyalkylamino. An independent claim is also included for dyeing keratinic fibers by applying (VI) or (VII) onto the wet or dry fibers in the presence of an oxidizing agent. [Image].
专利号:US-9181559-B2 优先权日:2009-09-15 标题 :Generation of high polyhydroxybutyrate producing oilseeds 发明人:PATTERSON NII; TANG JIHONG; HAN JIXIANG; TAVVA VENKATA; HERTIG ANDREW; ZHANG ZHIGANG; RAMSEIER THOMAS MARTIN; BOHMERT-TATAREV KAREN; PEOPLES OLIVER P; SNELL KRISTI D 权利人:PATTERSON NII; TANG JIHONG; HAN JIXIANG; TAVVA VENKATA; HERTIG ANDREW; ZHANG ZHIGANG; RAMSEIER THOMAS MARTIN; BOHMERT-TATAREV KAREN; PEOPLES OLIVER P; SNELL KRISTI D; METABOLIX INC 摘要:Transgenic plants, plant material, plant cells, and genetic constructs for synthesis of biopolymers, for example polyhydroxyalkanoates (“PHAâ€?) are provided. In one embodiment, the transgenic plants synthesize polyhydroxybutyrate (“PHBâ€?). In one embodiment the transgenic plant encodes siRNA for one or more of the genes encoding enzymes for producing PHA. In a more preferred embodiment, the siRNA expression is under the control of an inducible regulatory element. In another embodiment, the transgenic plant contains transgenes that encode expression enzymes that will degrade the polymer. In a preferred embodiment, the expression of these enzymes is under the control of a germination specific, inducible, or minimal promoter. In another embodiment, the transgenic plant contains transgenes encoding enzymes that increase carbon flow for polymer synthesis. In a preferred embodiment, these transgenes encode enzymes that increase carbon flow in the Calvin Cycle.
专利号:US-7288120-B2 优先权日:2003-12-12 标题 :Synthesis of tetraazapentamethine compounds, and processes for dyeing keratin fibers using this synthesis 发明人:PEREIRA RUI; BURGAUD HERVE 权利人:OREAL 摘要:The present disclosure relates to processes for synthesizing tetraazapentamethine compounds that comprise reacting at least one compound of azine type with at least one oxidizing agent. The disclosure also relates to processes for dyeing keratin fibers, for example human keratin fibers, that comprise using such synthetic processes.
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