公开(公告)号：US20160168187A1

公开(公告)日：2016-06-16

申请号：US14910203

申请日：2014-08-08

Applicant: THE SCRIPPS RESEARCH INSTITUTE

Inventor： Floyd E. ROMESBERG ,  Denis A. MALYSHEV ,  Lingjun LI ,  Thomas LAVERGNE ,  Zhengtao LI

IPC: C07H19/24 ,  C12P19/34

CPC classification number: C07H19/24 ,  C07H19/00 ,  C07H21/00 ,  C12P19/34 ,  C12Q1/6832 ,  C12Q2525/117

Abstract: Provided herein are analogs of unnatural nucleotides bearing predominantly hydrophobic nucleobase analogs that form unnatural base pairs during DNA polymerase-mediated replication of DNA or RNA polymerase-mediated transcription of RNA. In this manner, the unnatural nucleobases can be introduced in a site-specific way into oligonucleotides (single or double stranded DNA or RNA), where they can provide for site-specific cleavage, or can provide a reactive linker than can undergo functionalization with a cargo-bearing reagent by means of reaction with a primary amino group or by means of click chemistry with an alkyne group of the unnatural nucleobase linker.

Abstract translation: 本文提供了在DNA聚合酶介导的DNA或RNA聚合酶介导的RNA转录的DNA聚合酶介导的复制期间，主要具有疏水性核碱基类似物的类似物的类似物，其形成非天然碱基对。 以这种方式，可以以位点特异性方式将非天然核碱基引入寡核苷酸（单链或双链DNA或RNA），其中它们可以提供位点特异性切割，或者可以提供比经过官能化的反应性连接体 通过与伯氨基反应或通过使用非天然核碱基连接体的炔基进行点击化学反应的载货试剂。

公开(公告)号：US20240218027A1

公开(公告)日：2024-07-04

申请号：US18503341

申请日：2023-11-07

Applicant: THE SCRIPPS RESEARCH INSTITUTE

Inventor： Floyd E. ROMESBERG ,  Yorke ZHANG

IPC: C07K14/405 ,  C12N9/22 ,  C12N15/10 ,  C12N15/11 ,  C12N15/90

CPC classification number: C07K14/405 ,  C12N9/22 ,  C12N15/10 ,  C12N15/11 ,  C12N15/90 ,  C12N2310/20 ,  C12N2800/80

Abstract: Disclosed herein are proteins, methods, cells, engineered microorganisms, and kits for generating a modified nucleoside triphosphate transporter from Phaeodactylum tricornutum. Also disclosed herein proteins, methods, cells, engineered microorganisms, and kits for production of a nucleic acid molecule that comprises an unnatural nucleotide utilizing a modified nucleoside triphosphate transporter from Phaeodactylum tricornutum.

公开(公告)号：US20220243244A1

公开(公告)日：2022-08-04

申请号：US17716848

申请日：2022-04-08

Applicant: The Scripps Research Institute

Inventor： Floyd E. ROMESBERG ,  Emil C. FISCHER ,  Koji HASHIMOTO ,  Aaron W. FELDMAN ,  Vivian T. DIEN ,  Yorke ZHANG

IPC: C12P21/02

Abstract: Disclosed herein are compositions, methods, and kits for a cell incorporating unnatural amino acids into an unnatural polypeptide. Also disclosed herein are compositions, methods, and kits for increasing activity and yield of the unnatural polypeptide synthesized by the cell.

公开(公告)号：US20200095591A1

公开(公告)日：2020-03-26

申请号：US16591422

申请日：2019-10-02

Applicant: The Scripps Research Institute

Inventor： Floyd E. ROMESBERG ,  Denis A. MALYSHEV

IPC: C12N15/70 ,  C07K14/405 ,  C07K14/295 ,  C12N15/82 ,  C12P19/34

Abstract: A recombinantly expressed nucleotide triphosphate transporter efficiently imports the triphosphates of unnatural nucleotides into cells, and the endogenous cellular machinery incorporates those nucleotides into cellular nucleic acids. UBPs can therefore form within the cell's nucleic acids. Moreover, neither the presence of the unnatural triphosphates nor the replication of the UBP represents a significant growth burden. The UBP is not efficiently excised by nucleic acid repair pathways, and therefore can be retained as long as the unnatural triphosphates are available in the growth medium. Thus, the resulting cell is the first organism to stably propagate an expanded genetic alphabet.

公开(公告)号：US20200040027A1

公开(公告)日：2020-02-06

申请号：US16518715

申请日：2019-07-22

Applicant: The Scripps Research Institute

Inventor： Floyd E. ROMESBERG ,  Denis A. MALYSHEV ,  Lingjun LI ,  Thomas LAVERGNE ,  Zhengtao LI

IPC: C07H19/24 ,  C07H19/00 ,  C07H21/00 ,  C12Q1/6832 ,  C12P19/34

Abstract: Provided herein are analogs of unnatural nucleotides bearing predominantly hydrophobic nucleobase analogs that form unnatural base pairs during DNA polymerase-mediated replication of DNA or RNA polymerase-mediated transcription of RNA. In this manner, the unnatural nucleobases can be introduced in a site-specific way into oligonucleotides (single or double stranded DNA or RNA), where they can provide for site-specific cleavage, or can provide a reactive linker than can undergo functionalization with a cargo-bearing reagent by means of reaction with a primary amino group or by means of click chemistry with an alkyne group of the unnatural nucleobase linker.

公开(公告)号：US20170029829A1

公开(公告)日：2017-02-02

申请号：US15302874

申请日：2015-04-09

Applicant: The Scripps Research Institute

Inventor： Floyd E. ROMESBERG ,  Denis A. MALYSHEV

IPC: C12N15/70 ,  C07K14/295 ,  C07K14/405

Abstract: A recombinantly expressed nucleotide triphosphate transporter efficiently imports the triphosphates of unnatural nucleotides into cells, and the endogenous cellular machinery incorporates those nucleotides into cellular nucleic acids. UBPs can therefore form within the cell's nucleic acids. Moreover, neither the presence of the unnatural triphosphates nor the replication of the UBP represents a significant growth burden. The UBP is not efficiently excised by nucleic acid repair pathways, and therefore can be retained as long as the unnatural triphosphates are available in the growth medium. Thus, the resulting cell is the first organism to stably propagate an expanded genetic alphabet.

Abstract translation: 重组表达的三磷酸核苷酸转运蛋白有效地将非天然核苷酸的三磷酸进入细胞，并且内源细胞机制将这些核苷酸掺入到细胞核酸中。 因此，UBPs可以在细胞的核酸内形成。 此外，不自发三磷酸的存在和UBP的复制都不会代表显着的生长负担。 UBP没有被核酸修复途径有效地切除，因此只要在生长培养基中存在非天然三磷酸，就可以保留UBP。 因此，所得细胞是稳定繁殖扩展的遗传字母表的第一生物体。

公开(公告)号：US20250066799A1

公开(公告)日：2025-02-27

申请号：US18947037

申请日：2024-11-14

Applicant: THE SCRIPPS RESEARCH INSTITUTE

Inventor： Floyd E. ROMESBERG ,  Michael P. LEDBETTER ,  Rebekah J. KARADEEMA

IPC: C12N15/67 ,  C07K14/405 ,  C12N9/22 ,  C12N9/96 ,  C12N15/113 ,  C12N15/70 ,  C12P19/34

Abstract: Disclosed herein are methods, cells, engineered microorganisms, and kits for increasing the production of polypeptides comprising one or more unnatural amino acids. Further provided are cells, engineered microorganisms, and kits for increasing the retention of unnatural nucleic acids encoding the unnatural amino acids in an engineered cell, or semi-synthetic organism.

公开(公告)号：US20240294962A1

公开(公告)日：2024-09-05

申请号：US18532609

申请日：2023-12-07

Applicant: The Scripps Research Institute

Inventor： Floyd E. ROMESBERG ,  Vivian T. DIEN ,  Aaron W. FELDMAN ,  Rebekah J. KARADEEMA ,  Lingjun LI ,  Michael P. LEDBETTER ,  Anne Xiaozhou ZHOU

IPC: C12P19/34 ,  C07H19/24 ,  C07H21/04

CPC classification number: C12P19/34 ,  C07H19/24 ,  C07H21/04

Abstract: Disclosed herein are compositions, methods, cells, engineered microorganisms, and kits for increasing the production of proteins or polypeptides comprising one or more unnatural amino acids. Further provided are compositions, cells, engineered microorganisms, and kits for increasing the retention of unnatural nucleic acids encoding the unnatural amino acids in an engineered cell, or semi-synthetic organism.

公开(公告)号：US20240101590A1

公开(公告)日：2024-03-28

申请号：US18112952

申请日：2023-02-22

Applicant: The Scripps Research Institute

Inventor： Floyd E. ROMESBERG ,  Denis A. MALYSHEV ,  Lingjun LI ,  Thomas LAVERGNE ,  Zhengtao LI

IPC: C07H19/24 ,  C07H19/00 ,  C07H21/00 ,  C07H21/04 ,  C12N9/22 ,  C12N15/10 ,  C12P19/34 ,  C12Q1/6832

CPC classification number: C07H19/24 ,  C07H19/00 ,  C07H21/00 ,  C07H21/04 ,  C12N9/22 ,  C12N15/1024 ,  C12P19/34 ,  C12Q1/6832

Abstract: Provided herein are analogs of unnatural nucleotides bearing predominantly hydrophobic nucleobase analogs that form unnatural base pairs during DNA polymerase-mediated replication of DNA or RNA polymerase-mediated transcription of RNA. In this manner, the unnatural nucleobases can be introduced in a site-specific way into oligonucleotides (single or double stranded DNA or RNA), where they can provide for site-specific cleavage, or can provide a reactive linker than can undergo functionalization with a cargo-bearing reagent by means of reaction with a primary amino group or by means of click chemistry with an alkyne group of the unnatural nucleobase linker.

公开(公告)号：US20230392140A1

公开(公告)日：2023-12-07

申请号：US18304993

申请日：2023-04-21

Applicant: Synthorx, Inc. ,  The Scripps Research Institute

Inventor： Floyd E. ROMESBERG ,  Xiyu Dong ,  Anne Xiaozhou Zhou

IPC: C12N15/10 ,  C12N9/12

CPC classification number: C12N15/1048 ,  C12N9/1276 ,  C12Y207/07049 ,  C12N15/1096

Abstract: Disclosed herein are methods of reverse transcribing a polynucleotide comprising an unnatural ribonucleotide comprising reverse transcribing the polynucleotide with a reverse transcriptase in the presence of an unnatural dNTP comprising an unnatural nucleobase, wherein the reverse transcriptase polymerizes cDNA into which the unnatural NTP is incorporated. In some embodiments, the polynucleotide is present at a concentration less than or equal to about 500 nM and/or the polynucleotide is a tRNA, mRNA, RNA aptamer, or a member of a plurality of RNA aptamer candidates.