Abstract: Described herein is a method for editing the MHY7 gene in a cell by genome editing comprising introducing into the cell one or more deoxyribonucleic acid (DNA) endonucleases to effect one or more double stranded breaks (DSBs) within or near enhancer regions of the MYH7 gene or MYH6 gene that results in deletion of one or more enhancer regions of the MYH7 gene.
Type:
Application
Filed:
December 3, 2021
Publication date:
January 25, 2024
Inventors:
Elizabeth M. McNally, Anthony Martin Gacita
Abstract: The present disclosure provides RNA-guided CRISPR-Cas effector proteins, nucleic acids encoding same, and compositions comprising same. The present disclosure provides ribonucleoprotein complexes comprising: an RNA-guided CRISPR-Cas effector protein of the present disclosure; and a guide RNA. The present disclosure provides methods of modifying a target nucleic acid, using an RNA-guided CRISPR-Cas effector protein of the present disclosure and a guide RNA. The present disclosure provides methods of modulating transcription of a target nucleic acid.
Type:
Application
Filed:
May 8, 2023
Publication date:
January 25, 2024
Inventors:
Jennifer A. Doudna, Basem Al-Shayeb, Jillian F. Banfield, Patrick Pausch
Abstract: The present disclosure provides novel nucleic acid-guided nucleases and methods of using the nucleases for genome editing. The present disclosure further provides a system for editing a target region in a genome comprising a nucleic acid-guided nuclease, a heterologous guide nucleic acid for complexing with the nucleic acid-guided nuclease, and an editing polynucleotide configured to bind to the target region.
Type:
Application
Filed:
June 16, 2023
Publication date:
January 25, 2024
Inventors:
David Scott JOHNSON, Jan Fredrik SIMONS, Yoong Wearn LIM, Matthew James SPINDLER, Kyle Pierce CARTER, Savreet Kaur SANDHU, Ellen Kathleen WAGNER, Garry Coles, Robert EDGAR
Abstract: The present disclosure provides methods of modifying a target RNA in a eukaryotic cell. The present disclosure provides methods detecting a target RNA in a eukaryotic cell.
Abstract: The disclosure provides, e.g., compositions, systems, and methods for targeting, editing, modifying, or manipulating a host cell's genome at one or more locations in a DNA sequence in a cell, tissue, or subject.
Type:
Application
Filed:
August 10, 2023
Publication date:
January 25, 2024
Inventors:
Robert Charles Altshuler, Anne Helen Bothmer, Cecilia Giovanna Silvia Cotta-Ramusino, Randi Michelle Kotlar, Ananya Ray, Nathaniel Roquet, Carlos Sanchez, Barrett Ethan Steinberg
Abstract: The present disclosure provides engineered eukaryotic cells comprising a surface displayed catalytic domain of an endoglycosidase and methods of use.
Abstract: The present invention provides engineered amylase polypeptides and compositions thereof. The engineered amylase polypeptides have been optimized to provide improved thermostability, protease stability, and stability under a range of pH conditions, including acidic (pH <7) conditions. The invention also relates to the use of the compositions comprising the engineered amylase polypeptides for therapeutic and/or nutritional purposes. The present invention also provides polynucleotides encoding the engineered amylase polypeptides, as well as methods for making the engineered polynucleotides and amylase polypeptides.
Type:
Application
Filed:
September 6, 2023
Publication date:
January 25, 2024
Inventors:
Chinping Chng, Nikki Dellas, Da Duan, Ravi David Garcia, Harvinder Chagger Maniar
Abstract: Modified PH20 hyaluronidase polypeptides, including modified polypeptides that exhibit increased stability and/or increased activity, are provided. Also provided are compositions and formulations and uses thereof.
Type:
Application
Filed:
June 22, 2023
Publication date:
January 25, 2024
Inventors:
Ge WEI, H. Michael Shepard, Qiping Zhao, Robert James Connor
Abstract: Modified PH20 hyaluronidase polypeptides, including modified polypeptides that exhibit increased stability and/or increased activity, are provided. Also provided are compositions and formulations and uses thereof.
Type:
Application
Filed:
June 23, 2023
Publication date:
January 25, 2024
Inventors:
Ge WEI, H. Michael Shepard, Qiping Zhao, Robert James Connor
Abstract: Provided herein are fusion protein comprising: an effector domain comprising a catalytic domain of a deubiquitinase, or a functional fragment or functional variant thereof; and a targeting domain comprising a moiety that specifically binds a nuclear protein. Also provided herein are methods of using the fusion proteins to treat a disease, including genetic diseases.
Abstract: Provided herein are fusion protein comprising: an effector domain comprising a catalytic domain of a deubiquitinase, or a functional fragment or functional variant thereof; and a targeting domain comprising a moiety that specifically binds a mitochondrial protein. Also provided herein are methods of using the fusion proteins to treat a disease, including genetic diseases.
Abstract: The present disclosure relates to methods of expressing recombinant neprosin polypeptide, host cells and expression vectors for expressing the recombinant neprosin polypeptide, and methods of using the expressed neprosin polypeptide for therapeutic and other purposes.
Type:
Application
Filed:
May 10, 2023
Publication date:
January 25, 2024
Inventors:
Arianna I. Celis Luna, Da Duan, Paul Campion Jordan, Kathleen Susan Molnar, Adam P. Silverman, Tricia Windgassen
Abstract: The present invention provides a liquid enzyme preparation of a protein deamidase having excellent activity stability. This liquid enzyme preparation that contains a protein deamidase and at least 30 w/v % of sorbitol and that has a pH of 5.5 or more can enhance the stability of protein deamidase activity.
Abstract: A polymer brush composed of statistical copolymers of hydrophilic and hydrophobic monomers for enzyme immobilization. The heterogeneous polymer brushes stabilized four different lipases against high temperatures. Notably, the statistical copolymers stabilized the four lipases to a greater extent than a homopolymer brush. Additionally, in the case of Rhizomucor miehei lipase, statistical copolymers stabilized the enzyme to a greater extent than homopolymers of either hydrophilic or hydrophobic monomers. The resulting increase in high-temperature stability enabled significant improvements in catalytic rates by operating reactions at elevated temperatures, which is desirable for enzyme catalysis and sensing applications. Additionally, stabilization against elevated temperatures implies stabilization against non-aqueous solvents, which is of critical importance to numerous applications of biocatalysts.
Type:
Application
Filed:
November 15, 2021
Publication date:
January 25, 2024
Applicant:
The Regents of the University of Colorado, a body corporate
Inventors:
JAMES S. WELTZ, JOEL L. KAAR, DANIEL K. SCHWARTZ, HECTOR SANCHEZ-MORAN
Abstract: Disclosed herein are compositions comprising magnetoelectric composite materials and collagen, along with uses and kits thereof. Described herein is the use of DNA aptamer assemblies of varying DNA length, structure, and sequence to both bind to collagen and other proteins, to then act as a biocompatible, degradable, reversible, or permanent 3D crosslinkers between proteins, and to service as a biologically functional material when using the appropriate aptamer sequence. Therefore, disclosed herein are compositions comprising collagen fibers crosslinked with DNA aptamers. Also disclosed are devices and implants made from or coated with collagen fibers crosslinked with DNA aptamers. Also disclosed are methods of making collagen fibers. Also disclosed are kits for producing collagen fibers. Also disclosed herein are compositions DNA aptamers in a collagen fiber matrix that stabilizes the DNA aptamer.
Type:
Application
Filed:
September 10, 2021
Publication date:
January 25, 2024
Inventors:
Josephine ALLEN, Jennifer S. ANDREW, Noah D. FERSON, Bryan D. JAMES
Abstract: A method of fixating a tissue sample including: treating the tissue sample with a nano-tissue fixative solution to form a fixated tissue sample. A composition of nucleic acids in the fixated tissue sample is not altered. The nano-tissue fixative solution includes acetic acid, at least one alcohol, chloroform, titanium dioxide nanoparticles, zinc oxide nanoparticles, and silver nanoparticles.
Type:
Application
Filed:
July 20, 2022
Publication date:
January 25, 2024
Applicant:
Imam Abdulrahman Bin Faisal University
Inventors:
Khaled Fikry SALAMA, Amal Abdullah ALODAINI
Abstract: A nucleic acid extraction device for molecular testing comprises a film-flipping mechanism, a cap mechanism, a dispensing mechanism, an extraction plate transfer mechanism, and a nucleic acid extraction module. A sealing film is adhered onto top of an extraction plate and folded along an outer edge of the extraction plate. A film-flipping head of the film-flipping mechanism abuts against a bottom surface of the extraction plate and pushes an edge of the sealing film to unfold the sealing film. A film gripper clamps and removes the unfolded sealing film. The cap mechanism opens the specimen containers. The dispensing mechanism transfers the specimens from the specimen containers to the extraction plate. The extraction plate transfer mechanism moves the extraction plate with the specimens to the nucleic acid extraction module. As a result, molecular testing is automated to reduce labor and improve quality.
Abstract: A treatment apparatus includes a heating block (11) configured to heat a container (30) accommodating a sample (100) including a liquid within a first range (11A) in a gravity direction, and a cooling block (12) configured to cool the container (30) heated by the heating block (11) within a second range (12A) narrower than the first range (11A) in the gravity direction.
Abstract: A system, methods, and apparatus are described to collect and prepare single cells and groups of cells from microsamples of specimens and encode spatial information of the physical position of the cells in the specimen. In some embodiment, beads or surfaces with oligonucleotides containing spatial barcodes are used to analyze DNA or RNA. The spatial barcodes allow the position of the cell to be defined and the nucleic acid sequencing information, such as target sequencing, whole genome, gene expression, used to analyze the cells in a microsample for cell type, expression pattern, DNA sequence, and other information, in the context of the cell's physical position in the specimen. In other embodiment, markers such as isotopes are added to a microsample to encode spatial position with mass spectoscopy or other analysis. The spatial encoded information is then readout by analysis such as DNA sequencing, mass spectrometry, fluorescence, or other methods.
Abstract: The present invention relates to amphiphiles and specifically the amphiphiles that can be used as cell lysis reagents. The invention reveals the amphiphiles as well as their combinations for efficiently lysing the mammalian cells under relatively mild conditions that are compatible with a reverse transcription and polymerase chain reaction. The invention reveals the use of mixture of amphiphiles to improve the cell lysis and obtain increased yields of copy DNA during the reverse transcription reaction. The invention also provides a method for increasing the diversity of gene and transcript capture during single-cell RNA-sequencing using droplet microfluidics.
Abstract: Methods and devices for stabilizing a biological sample for analysis, comprising the steps of obtaining in a sample collection device a biological sample from a subject, the biological sample including at least one circulating cell-free first nucleic acid from the subject. The methods may include a step of contacting the biological sample while within the sample collection device with a protective agent composition that includes a preservative agent, an optional anticoagulant, and a quenching agent to form a mixture that includes the protective agent composition and the sample.
Type:
Application
Filed:
September 27, 2023
Publication date:
January 25, 2024
Inventors:
Wayne L. Ryan, M. Rohan Femando, Kausik Das
Abstract: The present disclosure provides methods and systems for purification of chemically modified, specifically, sulfonated DNA. Additionally, the disclosure provides methods and systems for bisulfite conversion with improved purification of sulfonated DNA using silica supports, such as silica beads, and acidic conditions for binding sulfonated DNA to silica surfaces.
Type:
Application
Filed:
July 29, 2021
Publication date:
January 25, 2024
Inventors:
Evan M. Ragland, Zubin Gagrat, Michael J. Domanico
Abstract: Provided are fusion proteins and related molecules useful for conducting base editing with reduced or no off-target mutations. The fusion protein may include a first fragment comprising a nucleobase deaminase or a catalytic domain thereof, a second fragment comprising a nucleobase deaminase inhibitor, and a protease cleavage site between the first fragment and the second fragment. Also provided are improved prime editing systems, including prime editing guide RNA with improved stability.
Type:
Application
Filed:
September 29, 2023
Publication date:
January 25, 2024
Inventors:
Jia Chen, Bei Yang, Li Yang, Xingxu Huang, Lijie Wang
Abstract: Provided is a quick and simple method for screening dimerized cyclic peptides, a library therefor, and molecules including cyclic peptides that can be used in the method and the library. For example, the method for screening dimerized cyclic peptides includes (1) a step in which, in a cell-free expression system, a cyclic peptide, a dimerization domain including a cysteine residue and a leucine zipper region, and at least one polypeptide including a peptide linker existing between the cyclic peptide and the dimerization domain are synthesized, and a dimer of the polypeptide is prepared, and (2) a step in which activity based on binding between the dimer and a target molecule is measured.
Abstract: The present disclosure relates to the field of affinity chromatography, and more specifically to the provision of nucleic acid and polypeptide libraries encoding three-helix bundle protein domains suitable for selecting affinity ligands that specifically binds to a target molecule of interest. The disclosure also relates to methods of using those libraries to identify and isolate such affinity ligands to a target molecule.
Abstract: Among the various aspects of the present disclosure is the provision of compositions for single-cell reporter assays and methods of use thereof. Also provided are methods of determining individual activities of a plurality of nucleic acid regulatory elements, identifying a regulatory element having cell type-specific activity, or determining variance in activity of a plurality of nucleic acid regulatory elements.
Abstract: Provided herein is an antigen display library for detecting antibodies produced by an individual; and methods of using the antigen display library to generate an antibody signature, the method comprising contacting a biological sample containing antibodies from an individual with the antigen display library, isolating phage clones displaying antigenic epitopes recognized by antibody in the sample, and identifying the antigenic epitopes that were recognized by antibody in the sample. Also provided are kits for generating an antibody signature comprising the antigen display library, a substrate for isolating phage clones bound by antibody, and may further comprise reagents useful for generating the antibody signature.
Abstract: The present disclosure discloses methods and systems for encoding digital information in nucleic acid (e.g., deoxyribonucleic acid) molecules without base-by-base synthesis, by encoding bit-value information in the presence or absence of unique nucleic acid sequences within a pool, comprising specifying each bit location in a bit-stream with a unique nucleic sequence and specifying the bit value at that location by the presence or absence of the corresponding unique nucleic acid sequence in the pool. Also disclosed are chemical methods for generating unique nucleic acid sequences using combinatorial genomic strategies (e.g., assembly of multiple nucleic acid sequences or enzymatic-based editing of nucleic acid sequences).
Type:
Application
Filed:
August 4, 2023
Publication date:
January 25, 2024
Inventors:
Devin Leake, Milena Lazova, Sarah Flickinger, Nathaniel Roquet, Hyunjun Park, Swapnil P. Bhatia
Abstract: Materials and methods for preparing nucleic acid libraries for next-generation sequencing are described herein. A variety of approaches are described relating to the use of unique molecular identifiers with transposon-based technology in the preparation of sequencing libraries. Also described herein are sequencing materials and methods for identifying and correcting amplification and sequencing errors.
Inventors:
Susan C. Verity, Robert Scott Kuersten, Niall Anthony Gormley, Andrew B. Kennedy, Sarah E. Shultzaberger, Andrew Slatter, Emma Bell, Sebastien Georg Gabriel Ricoult, Grace DeSantis, Fiona Kaper, Han-Yu Chuang, Oliver Jon Miller, Jason Richard Betley, Stephen M. Gross, Mats Ekstrand
Abstract: An improved method for Next Generation Sequencing which relies on the presence of the same distinct unique molecular identifier (UMI) located at each end of a linear nucleic acid molecule so that sequence reads of approximately 2 kb or longer are obtained, and which allows generation of a genomic map without the need of a reference sequence.
Abstract: Disclosed herein is a polynucleotide construct comprising one or more nuclease recognition sequences upstream and downstream of a Gene editing multi-site that comprises a plurality of recognition sequences for a site-specific recombinase or a nuclease. The plurality of recognition sequences facilitate insertion of one or more exogenous donor genes into the host cell.
Abstract: The present invention relates to compositions comprising RNA guides targeting TRAC, processes for characterizing the compositions, cells comprising the compositions, and methods of using the compositions.
Abstract: Disclosed herein are CRISPR/Cas systems comprising a fusion protein and at least one gRNA targeting a gene or a regulatory element thereof in a cell such as an immune cell, and vector compositions encoding the same. The systems and compositions may be used in methods of modulating expression of a gene in a cell such as an immune cell, as well as in methods of treating a disease such as cancer, autoimmune diseases, or viral infections.
Abstract: Provided are compounds, methods, and pharmaceutical compositions for reducing the amount or activity of SCN2A RNA in a cell or subject, and in certain instances reducing the amount of SCN2A protein in a cell or subject. Such compounds, methods, and pharmaceutical compositions are useful to ameliorate at least one symptom or hallmark of a disease or disorder associated with a voltage-gated sodium channel protein, such as, for example, a Developmental and Epileptic Encephalopathy, an intellectual disability, or an autism spectrum disorder. Such symptoms and hallmarks include, but are not limited to seizures, hypotonia, sensory integration disorders, motor development delays and dysfunctions, intellectual and cognitive dysfunctions, movement and balance dysfunctions, visual dysfunctions, delayed language and speech, gastrointestinal disorders, neurodevelopmental delays, sleep problems, and sudden unexpected death in epilepsy.
Type:
Application
Filed:
August 6, 2021
Publication date:
January 25, 2024
Inventors:
Paymaan Jafar-Nejad, Huynh-Hoa Bui, Susan M. Freier, Frank Rigo
Abstract: Embodiments of the disclosure include methods for generating skeletal muscle by targeting the Hippo pathway. In particular embodiments, an individual with a need for skeletal muscle generation is provided an effective amount of a shRNA molecule that targets the SAV1 gene. Particular shRNA sequences are disclosed.
Type:
Application
Filed:
November 18, 2021
Publication date:
January 25, 2024
Inventors:
Richard A.F. Dixon, Qi Liu, James T. Willerson, James F. Martin
Abstract: Disclosed herein are products, methods, and uses for treating, ameliorating, udaying the progression of, and/or preventing a muscular dystrophy or a cancer including, but not limited to, facioscapulohumeral muscular dystrophy (FSHD) or a sarcoma. More particularly, disclosed herein are RNA interference-based products, methods, and uses for inhibiting or downregulating the expression of double homeobox 4 (DUX4). Even more particularly, the disclosure provides nucleic acids comprising U7 DUX4 antisense sequences for inhibiting or downregulating the expression of DUX4 and methods of using said antisense sequences to inhibit or downregulate DUX4 expression in cells and/or in cells of a subject having a muscular dystrophy or a cancer including, but not limited to, FSHD or a cancer.
Type:
Application
Filed:
November 30, 2021
Publication date:
January 25, 2024
Inventors:
Scott Quenton Harper, Afrooz Rashnonejad, Nicolas Sebastien Wein
Abstract: A composition of matter comprising a synthetic miR-135 molecule comprising a nucleic acid sequence of a miR-135b as set forth in SEQ ID NO: 37, and a complementary strand as set forth in SEQ ID NO: 40 is disclosed. A conjugate comprising a composition of matter comprising a synthetic miR-135 molecule and a cell-targeting moiety is also disclosed.
Type:
Application
Filed:
July 14, 2023
Publication date:
January 25, 2024
Applicants:
Yeda Research and Development Co. Ltd., miCure Therapeutics Ltd.
Inventors:
Alon CHEN, Sharon MANASHIROV, Andres Pablo MONTEFELTRO
Abstract: Among other things, the present disclosure provides oligonucleotides and compositions thereof. In some embodiments, provided oligonucleotides and compositions are useful for adenosine modification. In some embodiments, the present disclosure provides methods for treating various conditions, disorders or diseases that can benefit from adenosine modification.
Type:
Application
Filed:
March 11, 2022
Publication date:
January 25, 2024
Inventors:
Prashant Monian, Chikdu Shakti Shivalila, Subramanian Marappan, Chandra Vargeese, Pachamuthu Kandasamy, Genliang Lu, Hui Yu, David Charles Donnell Butler, Luciano Henrique Apponi, Mamoru Shimizu, Stephany Michelle Standley, David John Boulay, Andrew Guzior Hoss, Jigar Desai, Jack David Godfrey, Hailin Yang, Naoki Iwamoto, Jayakanthan Kumarasamy, Anthony Lamattina, Tom Liantang Pu
Abstract: Provided herein are methods of treating ligament flavum hypertrophy and spinal stenosis including ligament flavum hypertrophy in a patient. The method comprises delivering to cells of hypertrophic ligament flavum in a patient a pharmaceutical composition comprising micro-RNA 29a or a precursor thereof, thereby decreasing type I and/or III collagen production by the cells. Alternatively, an antisense or RNAi reagent for knocking down type I collagen and/or type III collagen expression is delivered to cells of hypertrophic ligament flavum in a patient.
Type:
Application
Filed:
July 18, 2023
Publication date:
January 25, 2024
Inventors:
Joon Lee, Gwendolyn A. Sowa, Nam V. Vo, Richard Wawrose
Abstract: The present invention relates to antisense oligonucleotides that are capable of modulating expression of Tau in a target cell. The oligonucleotides hybridize to MAPT mRNA. The present invention further relates to conjugates of the oligonucleotide and pharmaceutical compositions and methods for treatment of Tauopathies, Alzheimer's disease, fronto-temporal dementia (FTD), FTDP-17, progressive supranuclear palsy (PSP), chronic traumatic encephalopathy (CTE), corticobasal ganglionic degeneration (CBD), epilepsy, Dravet syndrome, depression, seizure disorders and movement disorders.
Type:
Application
Filed:
July 21, 2023
Publication date:
January 25, 2024
Inventors:
Peter HAGEDORN, Anja Mølhart HØG, Richard E. OLSON, Marianne L. JENSEN
Abstract: Disclosed herein is a modified ribonucleotide comprising a nucleoside comprising N4-acetylcytidine and/or 5-hydroxymethyluridine, and polyribonucleotides comprising the same. Also provided herein are compositions comprising a polyribonucleotide of the present disclosure and methods of making and using the same.
Abstract: Provided are interfering RNAs (e.g., siRNAs) targeting SARS-CoV (e.g., the POL, Spike, Helicase, or Envelop gene thereof) and therapeutic uses thereof for inhibiting SARS-CoV infection and/or treating diseases associated with the infection (e.g., COVID-19).
Abstract: Described are RNAi agents, compositions that include RNAi agents, and methods for inhibition of a Superoxide Dismutase 1 (SOD1) gene. The SOD1 RNAi agents and RNAi agent conjugates disclosed herein inhibit the expression of an SOD1 gene. Pharmaceutical compositions that include one or more SOD1 RNAi agents, optionally with one or more additional therapeutics, are also described. Delivery of the described SOD1 RNAi agents to central nervous system (CNS) tissue, in vivo, provides for inhibition of SOD1 gene expression and a reduction in SOD1 activity, which can provide a therapeutic benefit to subjects, including human subjects, for the treatment of various diseases including amyotrophic lateral sclerosis (ALS.
Type:
Application
Filed:
June 14, 2023
Publication date:
January 25, 2024
Inventors:
Christine Esau, Ji Young Suk, Tao Pei, Anthony Nicholas, Xiaokai Li, Jeffrey Carlson
Abstract: Disclosed herein are compositions and compounds comprising modified oligonucleotides for modulating TMPRSS6 and modulating an iron accumulation disease, disorder and/or condition in an individual in need thereof. Iron accumulation diseases in an individual such as polycythemia, hemochromatosis or ?-thalassemia can be treated, ameliorated, delayed or prevented with the administration of antisense compounds targeted to TMPRSS6.
Type:
Application
Filed:
July 25, 2023
Publication date:
January 25, 2024
Applicant:
Ionis Pharmaceuticals, Inc.
Inventors:
Shuling Guo, Mariam Aghajan, Eric E. Swayze
Abstract: Methods and compositions using dnayzme-based GATA3 inhibitors for the treatment of insulin resistance and Type II diabetes, for the stimulation of adipogenesis in vivo, and for the promotion of fat redistribution are disclosed.
Abstract: A dual-specific aptamer comprising the nucleic acid sequence set forth in SEQ ID NO: 14. The dual-specific aptamer comprising an anti-HER2 segment capable of binding to a HER2 marker and an anti-CD16 segment capable of binding to a CD16 marker. The anti-HER2 segment comprises nucleotide 1 to nucleotide 42 of SEQ ID NO: 14, and the anti-CD16 segment comprises nucleotide 63 to nucleotide 103 of SEQ ID NO: 14. The dual-specific aptamer further comprises a nucleic acid linker comprising nucleotide 43 to nucleotide 62 of SEQ ID NO: 14 and is selected from the group consisting of a double-stranded nucleic acid linker or a single-stranded nucleic acid linker.
Abstract: The present disclosure relates to targeted genome editing in eukaryotic cells or organisms. More particularly, the present disclosure provides compositions for inducing targeted disruption of endogenous genes in eukaryotic cells. The composition may comprise a Cas9/guide RNA complex with a Cas9 protein to which a NLS is linked, and a guide RNA having a crRNA and a tracrRNA. The crRNA may comprise i) a portion to be hybridized with a portion of the tracrRNA, and ii) a portion complementary to a target DNA of the endogenous genes. In some embodiments, the Cas/guide RNA complex may be formed in vitro before being introduced into the eukaryotic cell.
Abstract: Disclosed herein, are methods of controlling expression of one or more genes of interest in one or more bacterial cells using a eukaryotic transcription factor, and inducible binary expression systems thereof.
Abstract: The present invention generally relates to the technical field of targeted modification of a nucleotide sequence of interest in the genome of a plant by means of a site-specific nuclease, wherein the modification and precision is assisted by specifically enhancing DNA polymerase theta (Pol ?) activity to improve genome editing (GE) efficiencies, preferably for increasing targeted insertion of a DNA insert. The invention describes a method for increasing the efficiency of targeted transgene insertion in a plant or a plant cell. Further provided are methods to modulate endogenous repair pathways with the aim to favor Pol ? activity in the context of GE. Further disclosed are suitable sequences and GE tools suitable in the methods of the invention as well as cells, tissues, organs or materials obtainable by the methods. Finally, an expression construct assembly for conducting the methods of the invention is disclosed.