公开(公告)号：US12297236B2

公开(公告)日：2025-05-13

申请号：US18480851

申请日：2023-10-04

Applicant: California Institute of Technology

Inventor： Michael J. Flynn ,  Michael B. Elowitz ,  Acacia Hori ,  Viviana Gradinaru

IPC: C07K14/005 ,  C12N15/86 ,  A61K38/00

Abstract: Disclosed herein include methods, compositions, and kits suitable for robust and tunable control of payload gene expression. Some embodiments provide rationally designed circuits, including miRNA-level and/or protein-level incoherent feed-forward loop circuits, that maintain the expression of a payload at an efficacious level. The circuit can comprise a promoter operably linked to a polynucleotide encoding a fusion protein comprising a payload protein, a protease, and one or more self-cleaving peptide sequences. The payload protein can comprise a degron and a cut site the protease is capable of cutting to expose the degron. The circuit can comprise a promoter operably linked to a polynucleotide comprising a payload gene, a silencer effector cassette, and one or more silencer effector binding sequences.

公开(公告)号：US20250019691A1

公开(公告)日：2025-01-16

申请号：US18757460

申请日：2024-06-27

Applicant: California Institute of Technology

Inventor： Richard F. Horns ,  Michael B. Elowitz ,  Joe A. Martinez

IPC: C12N15/10 ,  C07K14/005 ,  C12N9/22 ,  C12N15/11

Abstract: Disclosed herein include methods, compositions, and kits suitable for use in the measurement of the states of living cells across time and for use in the delivery of polyribonucleotides and circuits. There are provided, in some embodiments, RNA exporter proteins comprising an RNA-binding domain, a membrane-binding domain, and an interaction domain capable of nucleating self-assembly. Disclosed herein include polynucleotides encoding reporter RNA molecule(s) or cargo RNA molecule(s). In some embodiments, a plurality of RNA exporter proteins are capable of self-assembling into lipid-enveloped nanoparticles (LNs) secreted from a cell in which the RNA exporter proteins are expressed, thereby generating a population of LNs comprising exported reporter RNA molecule(s) or a fusogen and exported cargo RNA molecule(s). Disclosed herein include export modulator proteins capable of, e.g., enhancing or suppressing LN export.

公开(公告)号：US20240158449A1

公开(公告)日：2024-05-16

申请号：US18480851

申请日：2023-10-04

Applicant: California Institute of Technology

Inventor： Michael J. Flynn ,  Michael B. Elowitz ,  Acacia Hori ,  Viviana Gradinaru

IPC: C07K14/005 ,  C12N15/86

CPC classification number: C07K14/005 ,  C12N15/86 ,  A61K38/00

Abstract: Disclosed herein include methods, compositions, and kits suitable for robust and tunable control of payload gene expression. Some embodiments provide rationally designed circuits, including miRNA-level and/or protein-level incoherent feed-forward loop circuits, that maintain the expression of a payload at an efficacious level. The circuit can comprise a promoter operably linked to a polynucleotide encoding a fusion protein comprising a payload protein, a protease, and one or more self-cleaving peptide sequences. The payload protein can comprise a degron and a cut site the protease is capable of cutting to expose the degron. The circuit can comprise a promoter operably linked to a polynucleotide comprising a payload gene, a silencer effector cassette, and one or more silencer effector binding sequences.

公开(公告)号：US20250101424A1

公开(公告)日：2025-03-27

申请号：US18895366

申请日：2024-09-24

Applicant: CALIFORNIA INSTITUTE OF TECHNOLOGY

Inventor： Michael B. Elowitz ,  Rongrong Du ,  Michael J. Flynn

IPC: C12N15/113 ,  C12N15/85

Abstract: Disclosed herein include methods, compositions, and kits suitable for use in tuned dosage-invariant expression of a payload protein. Compositions (e.g., nucleic acid compositions, one or more cells) provided herein can comprise a first promoter sequence operably linked to a first polynucleotide comprising one or more miRNA cassettes, and a second promoter sequence operably linked to a second polynucleotide comprising a payload gene. In some embodiments, the first promoter sequence is capable of inducing transcription of the first polynucleotide to generate a first transcript, and the first transcript is capable of being processed to generate said miRNA. The payload gene can comprise a miRNA target region comprising one or more miRNA target sequences.

公开(公告)号：US11999964B2

公开(公告)日：2024-06-04

申请号：US17460120

申请日：2021-08-27

Applicant: California Institute of Technology

Inventor： Yitong Ma ,  Mark W. Budde ,  Michaelle N. Mayalu ,  Michael B. Elowitz

IPC: C12N15/85 ,  C07K14/415 ,  C12N9/02 ,  C12N9/86 ,  C12N15/11

CPC classification number: C12N15/85 ,  C07K14/415 ,  C12N9/0069 ,  C12N9/86 ,  C12N15/11 ,  C12Y113/12003 ,  C07K2319/00 ,  C12N2310/20 ,  C12N2800/107 ,  C12N2810/10

Abstract: Disclosed herein include circuits, compositions, nucleic acids, populations, systems, and methods enabling cells to sense, control, and/or respond to their own population size. In some embodiments, an orthogonal communication channel allows specific communication between engineered cells. Also described herein, in some embodiments, is an evolutionarily robust ‘paradoxical’ regulatory circuit architecture in which orthogonal signals both stimulate and inhibit net cell growth at different signal concentrations. In some embodiments, engineered cells autonomously reach designed densities and/or activate therapeutic or safety programs at specific density thresholds. Methods of treatment are also provided in some embodiments.

公开(公告)号：US11820794B2

公开(公告)日：2023-11-21

申请号：US17100857

申请日：2020-11-21

Applicant: California Institute of Technology

Inventor： Michael J. Flynn ,  Michael B. Elowitz ,  Acacia Hori ,  Viviana Gradinaru

IPC: C12N9/50 ,  A61K38/00 ,  C07K14/005 ,  C12N15/86

CPC classification number: C07K14/005 ,  C12N15/86 ,  A61K38/00 ,  C07K2319/71 ,  C07K2319/95

Abstract: Disclosed herein include methods, compositions, and kits suitable for robust and tunable control of payload gene expression. Some embodiments provide rationally designed circuits, including miRNA-level and/or protein-level incoherent feed-forward loop circuits, that maintain the expression of a payload at an efficacious level. The circuit can comprise a promoter operably linked to a polynucleotide encoding a fusion protein comprising a payload protein, a protease, and one or more self-cleaving peptide sequences. The payload protein can comprise a degron and a cut site the protease is capable of cutting to expose the degron. The circuit can comprise a promoter operably linked to a polynucleotide comprising a payload gene, a silencer effector cassette, and one or more silencer effector binding sequences.

公开(公告)号：US20230193317A1

公开(公告)日：2023-06-22

申请号：US17935024

申请日：2022-09-23

Applicant: California Institute of Technology

Inventor： Xiaojing Gao ,  Lucy S. Chong ,  Michael B. Elowitz ,  Matthew S-M Kim

IPC: C12N15/86 ,  C12N15/62 ,  C07K16/18 ,  C07K14/005 ,  C12N9/50 ,  C12N7/00

CPC classification number: C12N15/86 ,  C12N15/62 ,  C07K16/18 ,  C07K14/005 ,  C12Y304/21098 ,  C12N9/506 ,  C12N7/00 ,  C12N2820/007 ,  C07K2317/622 ,  C07K2317/569 ,  C07K2319/50 ,  C07K2319/30 ,  C12N2760/20143 ,  C12N2820/002 ,  C12N2740/11045 ,  C12N2810/6054

Abstract: Disclosed herein include methods, compositions, and systems suitable for use in delivering a polynucleotide to a target cell of a subject in need thereof. In some embodiments, a viral vector comprises a polynucleotide encoding nucleoprotein (N), phosphoprotein (P), matrix protein (M), RNA-dependent RNA polymerase (L), and one or more transgenes. The viral vector can comprise one or more of a conditionally stable fusion protein, a protease fusion protein, a degron fusion protein, and/or a glycoprotein derived of another species than the viral vector polynucleotide to enable control of viral vector transduction and/or replication.

公开(公告)号：US11421273B2

公开(公告)日：2022-08-23

申请号：US16701087

申请日：2019-12-02

Applicant: California Institute of Technology

Inventor： Amjad Askary ,  Michael B. Elowitz ,  Mark W. Budde ,  Carlos Lois ,  Luis Sanchez Guardado ,  Long Cai ,  James Linton

IPC: C12Q1/6874 ,  G01N1/30 ,  C12Q1/6876 ,  C12Q1/6818

Abstract: Disclosed herein include systems, methods, compositions, and kits for in situ readout of barcodes, such as DNA barcodes. Barcode constructs containing a promoter (e.g., a phage promoter) that is inactive in live cells can be integrated in the genomes of cells. Cells can be fixed, and phage RNA polymerase can be used for transcription of the barcode to RNA transcripts. The RNA transcripts can be detected using, for example, fluorescent imaging and used to determine barcode sequences.

公开(公告)号：US20200172968A1

公开(公告)日：2020-06-04

申请号：US16701087

申请日：2019-12-02

Applicant: California Institute of Technology

Inventor： Amjad Askary ,  Michael B. Elowitz ,  Mark W. Budde ,  Carlos Lois ,  Luis Sanchez Guardado ,  Long Cai ,  James Linton

IPC: C12Q1/6874 ,  C12Q1/6818 ,  C12Q1/6876 ,  G01N1/30

Abstract: Disclosed herein include systems, methods, compositions, and kits for in situ readout of barcodes, such as DNA barcodes. Barcode constructs containing a promoter (e.g., a phage promoter) that is inactive in live cells can be integrated in the genomes of cells. Cells can be fixed, and phage RNA polymerase can be used for transcription of the barcode to RNA transcripts. The RNA transcripts can be detected using, for example, fluorescent imaging and used to determine barcode sequences.

公开(公告)号：US20240101606A1

公开(公告)日：2024-03-28

申请号：US18316102

申请日：2023-05-11

Applicant: California Institute of Technology

Inventor： Lucy S. Chong ,  Ronghui Zhu ,  Xiaofei Ge ,  Michael B. Elowitz ,  Mark W. Budde

IPC: C07K14/00 ,  C12N9/22

CPC classification number: C07K14/001 ,  C12N9/22

Abstract: Disclosed herein include methods, compositions, and kits suitable for use in thresholding of protein signals. There are provided, in some embodiments, synthetic protein circuits that respond to inputs only above or below a certain threshold concentration. In some embodiments, the threshold value itself is tunable. Methods of treating a disease or disorder characterized by aberrant signaling are provided in some embodiments.