Tag: SpCas9

Crispr Library Download

CRISPR guide RNA libraries have been iteratively improved to provide increasingly efficient reagents, although their large size is a barrier for many applications. We design an optimised minimal genome-wide human CRISPR-Cas9 library (MinLibCas9) by mining existing large-scale gene loss-of-function datasets, resulting in a greater than 42% reduction in size compared…

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HUIADGENE ANNOUNCES RARE PEDIATRIC DRUG DESIGNATION GRANTED TO HG302, A NOVEL CRISPR DNA-EDITING THERAPY, FOR THE TREATMENT OF DUCHENNE MUSCULAR DYSTROPHY

HG302 effectively generates stable genome editing with minimal off-target effects in-vitro and in-vivo A single intravenous injection of HG302 restores functional dystrophin protein expression HG302 demonstrates therapeutic effects by improving muscle functions in humanized DMD mice SHANGHAI and CLINTON, N.J., Dec. 19, 2023 /PRNewswire/ — HuidaGene Therapeutics (辉大基因”HuidaGene”) today announced…

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Computational analysis of Cas proteins unlocks new potential in HIV-1 targeted gene therapy

1Department of Microbiology and Immunology, College of Medicine, Drexel University, United States 2Institute for Molecular Medicine and Infectious Disease, College of Medicine, Drexel University, United States 3Sidney Kimmel Cancer Center, United States The final, formatted version of the article will be published…

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CRISPR’s Reach Widens: Enzyme Targets Nearly All Human Genes

A team of engineers at Duke University have developed a method to broaden the reach of CRISPR technologies. While the original CRISPR system could only target 12.5% of the human genome, the new method expands access to nearly every gene to potentially target and treat a broader range of diseases…

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New enzyme allows CRISPR technologies to accurately target almost all human genes

Credit: Nature Chemical Biology (2023). DOI: 10.1038/s41589-023-01481-5 A team of engineers at Duke University have developed a method to broaden the reach of CRISPR technologies. While the original CRISPR system could only target 12.5% of the human genome, the new method expands access to nearly every gene to potentially target…

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Precise base editing without unintended indels in human cells and mouse primary myoblasts

Base editors cause unintended indels at the target sites Several types of evolved base editors based on the CRISPR system have been developed for more accurate and efficient genome engineering21,37. Among these, AncBE4max and ABEmax were evolved by modifying codon usage, NLSs, and ancestral deaminase reconstructions21. These modifications greatly improve…

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IJMS | Free Full-Text | CRISPR/Cas9-Mediated Genome Editing in Cancer Therapy

1. Introduction Recently, the morbidity and mortality rates of cancer have been increasing rapidly, posing a significant threat to human health. Cancer, a refractory and multifaceted disease, fundamentally originates from a cumulative series of mutations in the cellular genome and epigenome. These mutations activate oncogenes and deactivate tumor suppressors, leading…

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Transcriptional and epigenetic regulators of human CD8+ T cell function identified through orthogonal CRISPR screens

Developing an epigenetic screening platform in human T cells Staphylococcus aureus Cas9 (SaCas9) has been extensively used for genome editing in vivo as its compact size (3,159 bp) relative to the conventional Streptococcus pyogenes Cas9 (SpCas9) enables packaging into adeno-associated virus26,27,28. However, SaCas9 has not been widely used for targeted gene…

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Engineering a compact genome-editing tool

RESEARCH HIGHLIGHT 26 October 2023 CRISPR–Cas systems incorporating Streptococcus pyogenes Cas9 (SpCas9) and Acidaminococcus sp. Cas12a (AsCas12a) are commonly used genome-editing tools in human cells. However, these Cas enzymes are at the size limit for packaging in adeno-associated virus (AAV)-based delivery vehicles, which has hampered their applications. Writing in Cell,…

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Amerigo Scientific Unveils CRISPR/Cas Enzymes to Expand

Amerigo Scientific, as a distributor focused on providing critical products and services to biomedical and life science communities, announced the release of a groundbreaking selection of CRISPR/Cas enzymes to its extensive life science portfolio, aimed at further expanding the vast potential of the CRISPR toolbox. This addition brings cutting-edge genetic…

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Compact Gene-Editing Enzyme Could Enable More Effective Clinical Therapies

Scientists headed by a team at the University of Tokyo have developed a new CRISPR-based gene-editing tool that they suggest could lead to better treatments for patients with genetic disorders. The tool is a version of the compact AsCas12f enzyme that incorporates mutations giving it the same effectiveness as the…

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Aldevron, Jennifer Doudna, and Innovative Genomics Institute Collaborate on In Vivo CRISPR Therapies for Neurologic Diseases

Resulting paper published in Molecular Therapy FARGO, N.D., Aug. 29, 2023 /PRNewswire/ — Aldevron, a leading global manufacturer of DNA, RNA and proteins used in cell and gene therapies and vaccine development, contributed to a publication in collaboration with 2020 Nobel Laureate Jennifer Doudna and the Innovative Genomics Institute (IGI)…

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Mycobacterial DNA-binding protein 1 is critical for BCG survival in stressful environments and simultaneously regulates gene expression

Koul, A. et al. Delayed bactericidal response of Mycobacterium tuberculosis to bedaquiline involves remodelling of bacterial metabolism. Nat. Commun. doi.org/10.1038/ncomms4369 (2014). Article  PubMed  Google Scholar  Global Tuberculosis Report 2021. www.who.int/publications/digital/global-tuberculosis-report-2021. Colditz, G. A. et al. Efficacy of BCG vaccine in the prevention of tuberculosis: Meta-analysis of the published literature. JAMA…

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Optimized minimal genome-wide human sgRNA library

sgRNA collection and enzyme cut site annotation All sgRNAs corresponding to potential NGG-containing target sites on the human (GRCh38/hg38) genome were calculated by a customized Perl script. The enzyme cut site was set at the 17 position from 5’ to 3’ of each sgRNA sequence. These sgRNAs have more than…

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Custom CRISPR sgRNA Lentiviral Vectors & Viruses

Use abm’s All-in-One lentivectors and lentiviruses that express spCas9 or FnCas12a (FnCpf1) along with the sgRNA to simplify the delivery of CRISPR/Cas9 technology right into the target cell. Alternatively, custom sgRNA lentivectors and lentiviruses can be used to deliver sgRNAs into cells that already express the spCas9 nuclease, and thus…

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HKUMed establishes the first platform to slas

image: A new one-shot method enables testing all barcode-labelled precise genome editors’ performance with ease, circumventing the need of laborious one-by-one tests. view more  Credit: The University of Hong Kong A research team from the LKS Faculty of Medicine, the University of Hong Kong (HKUMed) has developed a new way to…

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Cas9 Proteins | Applied Biological Materials Inc.

CRISPR is the most versatile technology for genome editing and abm offers the largest selection of Cas9 proteins for CRISPR experiments. Cas9 Ribonucleoproteins (RNPs) are Cas9 proteins in complex with in vitro transcribed sgRNA. These RNPs can be used for in vitro pre-screening sgRNA candidates and can be directly transfected…

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Base Editors and Prime Editors Begin to Realize Their Clinical Promise

By MaryAnn Labant Back in 2016, a group of researchers led by Harvard’s David Liu, PhD, published a paper in Nature that reported the development of base editing. Base editing, they wrote, is “a new approach to genome editing that enables the direct, irreversible conversion of one target DNA base…

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Lack of Cas13a inhibition by anti-CRISPR proteins from Leptotrichia prophages

Summary CRISPR systems are prokaryotic adaptive immune systems that use RNA-guided Cas nucleases to recognize and destroy foreign genetic elements. To overcome CRISPR immunity, bacteriophages have evolved diverse families of anti-CRISPR proteins (Acrs). Recently, Lin et al. (2020) described the discovery and characterization of 7 Acr families (AcrVIA1–7) that inhibit…

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Gates Center Stem Cell Biobank and Disease Modeling Core

Stem Cell Biobank and Disease Modeling Core Overview   The Stem Cell Biobank and Disease Modeling Core (SCB&DM core) offers complete services related to the production of high quality human induced pluripotent stem cells (iPSCs) from patient-derived somatic cells and genetic engineering of iPSCs.  The core employs a patent-pending, RNA-based methodology to…

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Metagenomi Swiftly Variegates Gene-Editing Palatte

Despite the rapid development of genetic medicines, Simon Harnest, chief investment officer and SVP strategy at Metagenomi, said that attending the annual ASGCT meeting is a good way to keep tabs on the industry. “There are so many specialized groups in gene editing, some that focus on base editing and…

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HuidaGene Therapeutics Novel Cas12i DNA Gene Editing System Has Been Patented by…

17.05.2023 – 14:17 Huidagene Therapeutics SHANGHAI and CLINTON, NJ, April 20 /PRNewswire/ HuidaGene Therapeutics (辉大基因; HuidaGene), a clinical-stage company focused on the development of gene editing tools and gene therapies, announced today that the United States Patent and Trademark Office (USPTO) has granted the Company has granted patent US11,649,444B1 for…

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RCSB PDB – 8G1I: SpCas9 with sgRNA and target DNA

Find similar nucleic acids by:  Sequence  100% 95% 90% 80% 70% 60% 50% 40% 30% (by identity cutoff)  |  3D Structure Entity ID: 1 Molecule Chains Length Organism Image sgRNA A [auth B] 98 Streptococcus pyogenes Protein Feature View Expand Reference Sequence Find similar nucleic acids by:  Sequence  100% 95%…

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HuidaGene Therapeutics’ Novel DNA Gene-Editing System Cas12i Patent Granted by USPTO

SHANGHAI and CLINTON, N.J., May 16, 2023 /PRNewswire/ — HuidaGene Therapeutics (辉大基因; HuidaGene), a clinical-stage company focusing on developing gene editing tools and gene therapies, today announced that the United States Patent and Trademark Office (USPTO) has granted to the Company patent US11,649,444B1 with respect to its independently-developed DNA editing system,…

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SygRNA Cas9 Synthetic TracrRNA | TRACRRNA05N-5NMOL | SIGMA ALDRICH

Application Functional Genomics/Transgenic Applications Creation of gene knockouts Creation of knock-in animals or cell lines with promoters, fusion tags or reporters integrated into endogenous genes Features and Benefits HPLC-purified SygRNA™ Cas9 tracrRNA oligo is optimized for use with a target-specific cRNA and a source of SpCas9 protein for genome editing…

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Image1_Editing efficiencies with Cas9 orthologs, Cas12a endonucleases, and temperature in rice.TIF

The advent of CRISPR-Cas technology has made it the genome editing tool of choice in all kingdoms of life, including plants, which can have large, highly duplicated genomes. As a result, finding adequate target sequences that meet the specificities of a given Cas nuclease on any gene of interest remains…

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sygrna – Bjavkoreannew

SygRNA Cas9 Synthetic Modified tracrRNA; HPLC-purified SygRNA modified tracrRNA oligo is optimized for use with a target-specific crRNA or modified crRNA …The SygRNA™ synthetic RNA system recruits SpCas9 nuclease to specific genomic targets. The tracrRNA (trans-activating crRNA) binds to a crRNA (CRISPR RNA) …16 nov. 2016 — SygRNA™-Synthetic Two Part CRISPR RNA…

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Nonspecific interactions between Cas12a and dsDNA located downstream of the PAM mediate target search and assist AsCas12a for DNA cleavage

Cas12a is one of the most commonly-used Cas proteins for genome editing and gene regulation. The first key step for Cas12a to fulfill its function is to search for its target among numerous nonspecific and off-target sites. Cas12a utilizes one-dimensional diffusion along the contour of dsDNA to efficiently search for…

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SpCas9 sgRNA stem loop (medium with buldge)

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Hypertrophic Cardiomyopathy Mutation Reversed in Human Cell Line, Mouse Model With Gene Editing

In Nature Medicine, a University of Texas Southwestern Medical Center team describes a gene editing approach for correcting a hypertrophic cardiomyopathy (HCM)-linked MYH7 beta-myosin-coding gene mutation in human cardiomyocyte cells generated from induced pluripotent stem cells (iPSCs) and in a mouse model of HCM. Using their adenine base editor (ABE)…

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5fw3 – Crystal structure of SpCas9 variant VRER bound to sgRNA and TGCG PAM target DNA – Downloads

File format File name (file size) PDBx/mmCIF 5fw3.cif.gz Display(948.59 KB)5fw3.cif PDBx/mmJSON all 5fw3.json.gz Display (Tree)(617.41 KB)5fw3.json no-atom 5fw3-noatom.json.gz Display (Header)(38.55 KB)5fw3-noatom.json add only 5fw3-plus.json.gz Display(1.09 KB)5fw3-plus.json PDBML all 5fw3.xml.gz Display(1.24 MB)5fw3.xml no-atom 5fw3-noatom.xml.gz Display(66.00 KB)5fw3-noatom.xml ext-atom 5fw3-extatom.xml.gz Display(559.76 KB)5fw3-extatom.xml PDB pdb5fw3.ent.gz Display(820.75 KB)pdb5fw3.ent RDF 5fw3.rdf.gz Visualize(176.35 KB)5fw3.rdf Structure factors r5fw3sf.ent.gz…

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The CRISPR/Cas9 syst…

The CRISPR/Cas9 system is made of Cas9 nuclease and single-guide RNA (sgRNA). The sgRNA is an engineered single RNA molecule containing crispr RNA and tracr RNA parts. The sgRNA recognizes the target sequence by standard Watson-Crick base pairing. It has to be followed by a DNA motif called a protospacer…

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Evolution of giant pandoravirus revealed by CRISPR/Cas9

Viral strains utilized in this work The following viral strains have been used in this study: Pandoravirus neocaledonia3, Pandoravirus macleodensis3, Pandoravirus kuranda (this study, ON887157), Mollivirus kamchatka26, Pithovirus sibericum27 and Mimivirus reunion28. Cloning of DNA constructs All primers used in this study are listed in Table S1. All vectors used in…

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Versatile and efficient genome editing with Neisseria cinerea Cas9

PAM identification of NcCas9 by PAM-DOSE The type II-C Cas9 orthologue NcCas9 is of small size (1082 aa) and is closely related to conventional Neisseria meningitidis Cas9 (NmeCas9)11 (Fig. 1a). The NcCas9 is 94% identical to Nme1Cas9, and the divergences lie mainly in the C-terminal PAM-interacting domain (PID) (Fig. 1a and S1)….

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Optimization of sgRNA expression strategy to generate multiplex gene-edited pigs

[1] Fan ZY, Liu ZG, Xu K, Wu TW, Ruan JX, Zheng XM, et al. 2022. Long-term, multidomain analyses to identify the breed and allelic effects in MSTN-edited pigs to overcome lameness and sustainably improve nutritional meat production. Science China Life Sciences, 65(2): 362−375. doi:…

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CRISPR-SpRYgests Enable Precise Cleavage of DNA Bases In Vitro

NEW YORK — A CRISPR-Cas variant engineered to no longer need a protospacer adjacent motif (PAM) can be harnessed to make cuts at any DNA base in vitro, a new study reports. Its ability to make precise breaks anywhere could be applied to a number of DNA engineering applications. Researchers…

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Dynamic mechanisms of CRISPR interference by Escherichia coli CRISPR-Cas3

In vitro reconstitution of Escherichia coli CRISPR-Cas3 interference E. coli CRISPR-Cas3 is generally well-characterized type I CRISPR complexes in vitro and in vivo32,33,37,38. However, recombinant EcoCas3 protein is difficult to purify because of poor solubility and propensity to aggregate at 37 °C25,26,30,39. Co-expression of HtpG chaperon40 and/or low temperature growth at…

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Sigma-Aldrich Files Substantive Preliminary Motion No. 1 to Deny Broad Priority Benefit to Its Earliest-filed Provisional Application | McDonnell Boehnen Hulbert & Berghoff LLP

On December 3rd, Senior Party Sigma-Aldrich filed its Substantive Preliminary Motion No. 1 in Interference No. 106,133 (which names the Broad Institute, Harvard University, and MIT (collectively, Broad) as Junior Party), asking the Patent Trial and Appeal Board to deny Broad benefit of its U.S. Provisional Application No. 61/736,527, filed…

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CRISPR-Cas9 Gene Therapy for Duchenne Muscular Dystrophy

Ishino Y, Shinagawa H, Makino K, et al. Nucleotide sequence of the iap gene, responsible for alkaline phosphatase isozyme conversion in Escherichia coli, and identification of the gene product. J Bacteriol. 1987;169:5429–33. CAS  PubMed  PubMed Central  Google Scholar  Jansen R, van Embden JDA, Gaastra W, et al. Identification of genes…

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Broad Files Substantive Preliminary Motion No. 3 to Designate Claims as not Corresponding to Count in Interference No. 106,133 | McDonnell Boehnen Hulbert & Berghoff LLP

On December 3rd, Junior Party the Broad Institute, Harvard University, and MIT (collectively, Broad) filed its Contingent Preliminary Motion No. 3 in Interference No. 106,133 (which names Sigma-Aldrich as Senior Party), asking the Patent Trial and Appeal Board to designate certain claims deemed in the Declaration as corresponding to the…

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AttCRISPR: a spacetime interpretable model for prediction of sgRNA on-target activity | BMC Bioinformatics

Dataset The dataset we used for training, validation and testing is the DeepHF dataset [17]. We extracted 55604, 58617, 56888 sgRNAs with activity (represented by insertion/deletion (indel)) for WT-SpCas9, eSpCas9(1.1) and SpCas9-HF1, respectively, from its source data, and use the same partition method to divide train set and test set….

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ToolGen Files Opposition to Broad Preliminary Motion No. 3 to De-Designate Claims as Corresponding to Either Interference Count | McDonnell Boehnen Hulbert & Berghoff LLP

On May 28th, Junior Party the Broad Institute, Harvard University, and MIT (collectively, “Broad”) filed its Substantive Preliminary Motion No. 3 in CRISPR Interference No. 106,126 (where ToolGen is the Senior Party).  This motion, pursuant to 37 C.F.R. §§ 41.121(a)(1)(iii) and 41.208(a)(1) requested that the Board de-designate Broad claims in these…

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Compact CasMINI CRISPR Tech Is Easier to Deliver to Cells, Could Have Broad Gene Therapy Potential

Scientists led by a team at Stanford University have developed a compact, efficient CRISPR-Cas system, called CasMINI, which is about half the size of existing CRISPR-Cas systems, and which could have broad utility for gene therapy applications as well as cell engineering. The researchers confirmed in experiments that CasMINI could,…

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CRISPR-Based Therapeutics Blaze an In Vivo Path to the Clinic

Therapeutic applications of genome editing were envisioned at least as early as the mid-1990s, when the first sequence-specific genome editing technologies emerged. Initially, such applications were considered distant prospects, but by 2012, they suddenly seemed near to hand. It was at that time that CRISPR technologies emerged. CRISPR, which stands…

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Industrializing CRISPR

Sponsored content brought to you by Kevin Holden, PhD Kevin Holden, PhD, Head of Science at Synthego, discusses the importance of industrializing CRISPR as the technology matures and makes inroads in the clinic. GEN: What’s new and interesting to you in the world of CRISPR? HOLDEN: Some of the most…

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CRISPR: Guide to gRNA design

Introduction to CRISPR in SnapGene Genome editing technology has been evolving for many years. The Holy Grail of genome engineering has always been to introduce a specific genetic change that affects only the genomic target and leaves no undesired changes in the DNA. The discovery and application of the bacterial…

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