67,221 results match your criteria hek293 cells

Gain-of-function mutation in ubiquitin-ligase KLHL24 causes desmin degradation and dilatation in hiPSC-derived engineered heart tissues.

J Clin Invest 2021 Jul 22. Epub 2021 Jul 22.

Department of Cardiology, University Medical Center Groningen, Groningen, Netherlands.

The start codon c.1A>G mutation in KLHL24, encoding ubiquitin-ligase KLHL24, results in the loss of 28 N-terminal amino acids (KLHL24-ΔN28) by skipping the initial start codon. In skin, KLHL24-ΔN28 leads to gain of function, excessively targeting intermediate filament keratin-14 for proteasomal degradation, ultimately causing epidermolysis bullosa simplex (EBS). Read More

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Up-regulation of gasdermin C in mouse small intestine is associated with lytic cell death in enterocytes in worm-induced type 2 immunity.

Proc Natl Acad Sci U S A 2021 Jul;118(30)

Monell Chemical Senses Center, Philadelphia, PA 19104;

"Taste-like" tuft cells in the intestine trigger type 2 immunity in response to worm infection. The secretion of interleukin-13 (IL-13) from type 2 innate lymphoid cells (ILC2) represents a key step in the tuft cell-ILC2 cell-intestinal epithelial cell circuit that drives the clearance of worms from the gut via type 2 immune responses. Hallmark features of type 2 responses include tissue remodeling, such as tuft and goblet cell expansion, and villus atrophy, yet it remains unclear if additional molecular changes in the gut epithelium facilitate the clearance of worms from the gut. Read More

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Trophoblast derived extracellular vesicles specifically alter the transcriptome of endometrial cells and may constitute a critical component of embryo-maternal communication.

Reprod Biol Endocrinol 2021 Jul 21;19(1):115. Epub 2021 Jul 21.

Department of Pathophysiology, Institute of Biomedicine and Translational Medicine, Faculty of Medicine, Tartu University, Tartu, Estonia.

Background: The period of time when the embryo and the endometrium undergo significant morphological alterations to facilitate a successful implantation-known as "window of implantation"-is a critical moment in human reproduction. Embryo and the endometrium communicate extensively during this period, and lipid bilayer bound nanoscale extracellular vesicles (EVs) are purported to be integral to this communication.

Methods: To investigate the nature of the EV-mediated embryo-maternal communication, we have supplemented trophoblast analogue spheroid (JAr) derived EVs to an endometrial analogue (RL 95-2) cell layer and characterized the transcriptomic alterations using RNA sequencing. Read More

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Cells Stimulated with More Than One Toll-Like Receptor-Ligand in the Presence of a MyD88 Inhibitor Augmented Interferon- via MyD88-Independent Signaling Pathway.

Viral Immunol 2021 Jul 20. Epub 2021 Jul 20.

Department of Bacteriology, United States Army Medical Research Institute of Infectious Diseases, Frederick, Maryland, USA.

Host exposure to pathogens engage multiple pathogen recognition receptors (PRRs) including toll-like receptors (TLRs); recruit intracellular signaling adaptor proteins primarily myeloid differentiation primary response protein 88 (MyD88) for activating downstream signaling cascades, which culminate in the production of type I interferons (IFNs), proinflammatory cytokines, and chemokines; and impede pathogen replication and dissemination. However, recent studies highlight that absence of MyD88 increased antiviral type I IFN induction, and MyD88 mice showed a higher survival rate compared with the low survival rate of the MyD88 mice, implicating MyD88 limits antiviral type I IFN response. As a single infectious agent may harbor multiple PRR agonists, which trigger different sets of TLR-initiated immune signaling, we examined whether MyD88 inhibition during stimulation of cells with more than one TLR-ligand would augment type I IFN. Read More

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Adhesion of Enteroaggregative Strains to HEK293 Cells.

Bio Protoc 2018 Apr 20;8(8):e2802. Epub 2018 Apr 20.

Department of Pediatrics, University of Virginia School of Medicine, Charlottesville, Virginia, United States of America.

Enteroaggregative (EAEC) is a recognized cause of acute diarrhea among both children and adults worldwide. EAEC strains are characterized by the presence of aggregative adherence fimbriae (AAF), which play a key role in pathogenesis by mediating attachment to the intestinal mucosa and by triggering host inflammatory responses. The aggregative adherence fimbria II (AAF/II) is the most important adherence factor of EAEC prototype strain 042 (EAEC042) to intestinal cells. Read More

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CAMK2/CaMKII activates MLKL in short-term starvation to facilitate autophagic flux.

Autophagy 2021 Jul 20:1-19. Epub 2021 Jul 20.

Department of Integrative Biology and Pharmacology, McGovern Medical School, the University of Texas Health Science Center at Houston, Houston, Texas, USA.

MLKL (mixed lineage kinase domain like pseudokinase) is a well-known core component of necrosome that executes necroptotic cell death upon phosphorylation by RIPK3 (receptor interacting serine/threonine kinase 3). Recent studies also implicate a role of MLKL in endosomal trafficking, which is not always dependent on RIPK3. Using mouse Neuro-2a and L929 as well as human HEK293 and HT29 cells, we show here that MLKL is phosphorylated in response to serum and amino acid deprivation from the culture medium, in a manner that depends on CAMK2/CaMKII (calcium/calmodulin dependent protein kinase II) but not RIPK3. Read More

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Characterization of a functional V vasopressin receptor in the male rat kidney, evidence for crosstalk between V and V receptor signaling pathways.

Am J Physiol Renal Physiol 2021 07 20. Epub 2021 Jul 20.

Institut de génomique fonctionnelle, University of Montpellier, Montpellier, France.

Although vasopressin V receptor (V-R) mRNA was detected in the kidney, the precise renal localization, pharmacological and physiological properties of this receptor remain unknown. Using the selective V agonist d[Leu, Lys]VP, either fluorescent or radioactive, we showed that V-R is mainly present in principal cells of the inner medullary collecting duct (IMCD) in the male rat kidney. Protein and mRNA expression of V-R were very low compared to V receptor (V-R). Read More

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Effects of Kumaizasa () Leaf Extract on Innate Immune Regulation in HEK293 Cells and Macrophages.

Anticancer Res 2021 Aug;41(8):4093-4100

Faculty of Pharmaceutical Sciences, Hokkaido University of Science, Sapporo, Japan

Background/aim: We investigated the effect of Kumaizasa leaf extract (KLE) on innate immunity using the HEK293 and RAW 264.7 cell lines.

Materials And Methods: KLE, lipopolysaccharides (LPS), or KLE with LPS were added to RAW 264. Read More

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Urease: Unsuspected Non-Enzymatic Properties Relevant to Pathogenicity.

Int J Mol Sci 2021 Jul 4;22(13). Epub 2021 Jul 4.

Laboratory of Neurotoxins, Brain Institute of Rio Grande do Sul (BRAINS) and School of Medicine, Pontifícia Universidade Católica do Rio Grande do Sul (PUCRS), Porto Alegre CEP 90610-000, RS, Brazil.

Infection by causes urinary stones and catheter incrustation due to ammonia formed by urease (PMU), one of its virulence factors. Non-enzymatic properties, such as pro-inflammatory and neurotoxic activities, were previously reported for distinct ureases, including that of the gastric pathogen . Here, PMU was assayed on isolated cells to evaluate its non-enzymatic properties. Read More

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Production of Adeno-Associated Virus Vectors in Cell Stacks for Preclinical Studies in Large Animal Models.

J Vis Exp 2021 Jun 30(172). Epub 2021 Jun 30.

Department of Pathobiology, University of Guelph;

Adeno-associated virus (AAV) vectors are among the most clinically advanced gene therapy vectors, with three AAV gene therapies approved for humans. Clinical advancement of novel applications for AAV involves transitioning from small animal models, such as mice, to larger animal models, including dogs, sheep, and nonhuman primates. One of the limitations of administering AAV to larger animals is the requirement for large quantities of high-titer virus. Read More

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8-Hydroxyquinolin-2(1H)-one analogues as potential β-agonists: Design, synthesis and activity study.

Eur J Med Chem 2021 Jul 10;224:113697. Epub 2021 Jul 10.

Department of Medicinal Chemistry, School of Pharmaceutical Engineering, Shenyang Pharmaceutical University, Shenyang, 110016, China; Key Laboratory of Structure-Based Drug Design and Discovery of Ministry of Education, Shenyang Pharmaceutical University, Shenyang, 110016, China. Electronic address:

β-Agonists that bind to plasmalemmal β-adrenoceptors causing cAMP accumulation are widely used as bronchodilators in chronic respiratory diseases. Here, we designed and synthesized a group of 8-hydroxyquinolin-2(1H)-one analogues and studied their β-agonistic activities with a cellular cAMP assay. Compounds B05 and C08 were identified as potent (EC < 20 pM) and selective β-agonists among the compounds tested. Read More

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Fed-batch production assessment of a tetravalent bispecific antibody: A case study on piggyBac stably transfected HEK293 cells.

N Biotechnol 2021 Jul 15;65:9-19. Epub 2021 Jul 15.

Department of Pharmaceutical Biosciences, Science for Life Laboratory, Uppsala University, Uppsala, Sweden. Electronic address:

The transition from preclinical biological drug development into clinical trials requires an efficient upscaling process. In this context, bispecific antibody drugs are particularly challenging due to their propensity to form aggregates and generally produce low titers. Here, the upscaling process for a tetravalent bispecific antibody expressed by a piggyBac transposon-mediated stable HEK293 cell pool has been evaluated. Read More

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Simple, fast and efficient iTOP-mediated delivery of CRISPR/Cas9 RNP in difficult-to-transduce human cells including primary T cells.

J Biotechnol 2021 Jul 13;338:71-80. Epub 2021 Jul 13.

NTrans Technologies BV, Heidelberglaan 7, 3584 CS Utrecht, the Netherlands. Electronic address:

The advent of the CRISPR/Cas9 system has transformed the field of human genome engineering and has created new perspectives in the development of innovative cell therapies. However, the absence of a simple, fast and efficient delivery method of CRISPR/Cas9 into primary human cells has been limiting the progress of CRISPR/Cas9-based therapies. Here, we describe an optimized protocol for iTOP-mediated delivery of CRISPR/Cas9 in various human cells, including primary T cells, induced pluripotent stem cells (hiPSCs), Jurkat, ARPE-19 and HEK293 cells. Read More

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In cardiac muscle cells, both adrenergic agonists and antagonists induce reactive oxygen species from NOX2 but mutually attenuate each other's effects.

Eur J Pharmacol 2021 Jul 13;908:174350. Epub 2021 Jul 13.

School of Life Sciences, Jawaharlal Nehru University, New Mehrauli Road, New Delhi, 110067, India. Electronic address:

In cardiac muscle cells adrenergic agonists stimulate the generation of reactive oxygen species, followed by redox signaling. We postulated that the antagonists would attenuate such reactive oxygen species generation by the agonists. H9c2 cardiac myoblasts, neonatal rat cardiac myocytes, and HEK293 cells expressing β/β adrenoceptors were stimulated with several agonists and antagonists. Read More

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A Single Xyloglucan Xylosyltransferase is Sufficient for Generation of the XXXG Xylosylation Pattern of Xyloglucan.

Plant Cell Physiol 2021 Jul 15. Epub 2021 Jul 15.

Department of Plant Biology, University of Georgia, Athens, GA 30602, USA.

Xyloglucan is the most abundant hemicellulose in the primary cell walls of dicots. Dicot xyloglucan is the XXXG-type consisting of repeating units of three consecutive xylosylated Glc residues followed by one unsubstituted Glc. Its xylosylation is catalyzed by xyloglucan 6-xylosyltransferases (XXTs) and there exist five XXTs (AtXXT1-5) in Arabidopsis. Read More

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Sesquiterpenes and sesquiterpenoids harbor modulatory allosteric potential and affect inhibitory GABA receptor function in vitro.

J Neurochem 2021 Jul 15. Epub 2021 Jul 15.

Institute for Clinical Neurobiology, University Hospital, Julius-Maximilians-Universität Würzburg, 97078, Würzburg, Germany.

Naturally occurring compounds such as sesquiterpenes and sesquiterpenoids (SQTs) have been shown to modulate GABA receptors (GABA Rs). In this study, the modulatory potential at GABA Rs of 11 SQTs was analyzed to characterize their potential neurotropic activity. Transfected HEK293 cells and primary hippocampal neurons were functionally investigated using electrophysiological whole-cell recordings. Read More

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Behavior of the Antibacterial Peptide Cyclo[RRRWFW], Explored Using a 3-Hydroxychromone-Derived Fluorescent Amino Acid.

Front Chem 2021 28;9:688446. Epub 2021 Jun 28.

Taras Shevchenko National University of Kyiv, Kyiv, Ukraine.

Labeling biomolecules with fluorescent labels is an established tool for structural, biochemical, and biophysical studies; however, it remains underused for small peptides. In this work, an amino acid bearing a 3-hydroxychromone fluorophore, 2-amino-3-(2-(furan-2-yl)-3-hydroxy-4-oxo-4H-chromen-6-yl)propanoic acid (FHC), was incorporated in a known hexameric antimicrobial peptide, cyclo[RRRWFW] (cWFW), in place of aromatic residues. Circular dichroism spectropolarimetry and antibacterial activity measurements demonstrated that the FHC residue perturbs the peptide structure depending on labeling position but does not modify the activity of cWFW significantly. Read More

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SPARK: A Transcriptional Assay for Recording Protein-Protein Interactions in a Defined Time Window.

Curr Protoc 2021 Jul;1(7):e190

Life Sciences Institute, University of Michigan, Ann Arbor, Michigan.

Protein-protein interactions (PPIs) are ubiquitously involved in cellular processes such as gene expression, enzymatic catalysis, and signal transduction. To study dynamic PPIs, real-time methods such as Förster resonance energy transfer and bioluminescence resonance energy transfer can provide high temporal resolution, but they only allow PPI detection in a limited area at a time and do not permit post-PPI analysis or manipulation of the cells. Integration methods such as the yeast two-hybrid system and split protein systems integrate PPI signals over time and allow subsequent analysis, but they lose information on dynamics. Read More

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Advancing a rapid, high throughput screening platform for optimization of lentivirus production.

Biotechnol J 2021 Jul 14:e2000621. Epub 2021 Jul 14.

Department of Chemical and Biological Engineering, and Center for Biotechnology & Interdisciplinary Studies, Rensselaer Polytechnic Institute, Troy, New York, USA.

Background: Lentiviral vectors (LVVs) hold great promise as delivery tools for gene therapy and chimeric antigen receptor T cell (CAR-T) therapy. Their ability to target difficult to transfect cells and deliver genetic payloads that integrate into the host genome makes them ideal delivery candidates. However, several challenges remain to be addressed before LVVs are more widely used as therapeutics including low viral vector concentrations and the absence of suitable scale-up methods for large-scale production. Read More

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Immunogenicity and Protective Efficacy of a Highly Thermotolerant, Trimeric SARS-CoV-2 Receptor Binding Domain Derivative.

ACS Infect Dis 2021 Jul 14. Epub 2021 Jul 14.

Australian Centre for Disease Preparedness (ACDP), Commonwealth Scientific and Industrial Research Organisation (CSIRO), 5 Portarlington Road, Geelong 3220, Victoria, Australia.

The receptor binding domain (RBD) of SARS-CoV-2 is the primary target of neutralizing antibodies. We designed a trimeric, highly thermotolerant glycan engineered RBD by fusion to a heterologous, poorly immunogenic disulfide linked trimerization domain derived from cartilage matrix protein. The protein expressed at a yield of ∼80-100 mg/L in transiently transfected Expi293 cells, as well as CHO and HEK293 stable cell lines and formed homogeneous disulfide-linked trimers. Read More

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Reprogrammed CRISPR-Cas13b suppresses SARS-CoV-2 replication and circumvents its mutational escape through mismatch tolerance.

Nat Commun 2021 07 13;12(1):4270. Epub 2021 Jul 13.

Cancer Immunology Program, Peter MacCallum Cancer Centre, Melbourne, Australia.

The recent dramatic appearance of variants of concern of SARS-coronavirus-2 (SARS-CoV-2) highlights the need for innovative approaches that simultaneously suppress viral replication and circumvent viral escape from host immunity and antiviral therapeutics. Here, we employ genome-wide computational prediction and single-nucleotide resolution screening to reprogram CRISPR-Cas13b against SARS-CoV-2 genomic and subgenomic RNAs. Reprogrammed Cas13b effectors targeting accessible regions of Spike and Nucleocapsid transcripts achieved >98% silencing efficiency in virus-free models. Read More

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Evaluating Peptides of and Their Inhibitory Potential against ACE, DPP-IV, and Oxidative Stress.

J Proteome Res 2021 Jul 13. Epub 2021 Jul 13.

Department of Biotechnology, CSIR-Institute of Himalayan Bioresource Technology, Palampur 176061, HP, India.

Royle ex Benth. is a high-altitude plant having great medicinal value. However, its medicinal value at the peptide level is still unknown, which limits its utility in the development of peptide-based therapeutics. Read More

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TSC2 regulates lysosome biogenesis via a non-canonical RAGC and TFEB-dependent mechanism.

Nat Commun 2021 07 12;12(1):4245. Epub 2021 Jul 12.

Pulmonary and Critical Care Medicine, Department of Medicine, Brigham and Women's Hospital, Harvard Medical School, Boston, MA, USA.

Tuberous Sclerosis Complex (TSC) is caused by TSC1 or TSC2 mutations, resulting in hyperactivation of the mechanistic target of rapamycin complex 1 (mTORC1). Transcription factor EB (TFEB), a master regulator of lysosome biogenesis, is negatively regulated by mTORC1 through a RAG GTPase-dependent phosphorylation. Here we show that lysosomal biogenesis is increased in TSC-associated renal tumors, pulmonary lymphangioleiomyomatosis, kidneys from Tsc2 mice, and TSC1/2-deficient cells via a TFEB-dependent mechanism. Read More

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Distinct pathways of homologous recombination controlled by the SWS1-SWSAP1-SPIDR complex.

Nat Commun 2021 07 12;12(1):4255. Epub 2021 Jul 12.

Developmental Biology Program, Memorial Sloan Kettering Cancer Center, New York, NY, USA.

Homology-directed repair (HDR), a critical DNA repair pathway in mammalian cells, is complex, leading to multiple outcomes with different impacts on genomic integrity. However, the factors that control these different outcomes are often not well understood. Here we show that SWS1-SWSAP1-SPIDR controls distinct types of HDR. Read More

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Protein kinase C-mediated phosphorylation and α2δ-1 interdependently regulate NMDA receptor trafficking and activity.

J Neurosci 2021 Jun 17. Epub 2021 Jun 17.

Department of Anesthesiology and Perioperative Medicine, The University of Texas MD Anderson Cancer Center, Houston, Texas 77030, USA

N-methyl-D-aspartate receptors (NMDARs) are important for synaptic plasticity associated with many physiological functions and neurological disorders. Protein kinase C (PKC) activation increases the phosphorylation and activity of NMDARs, and α2δ-1 is a critical NMDAR-interacting protein and controls synaptic trafficking of NMDARs. In this study, we determined the relative roles of PKC and α2δ-1 in the control of NMDAR activity. Read More

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A trivalent TNF-R2 as a new tumor necrosis factor alpha-blocking molecule.

Proteins 2021 Jul 11. Epub 2021 Jul 11.

Recombinant Biopharmaceuticals Laboratory, Pharmacology Department, School of Biological Sciences, Universidad de Concepción, Concepción, Chile.

The neutralization of tumor necrosis factor alpha (TNFα) with biopharmaceuticals is a successful therapy for inflammatory diseases. Currently, one of the main TNFα-antagonists is Etanercept, a dimeric TNF-R2 ectodomain. Considering that TNFα and its receptors are homotrimers, we proposed that a trimeric TNF-R2 ectodomain could be an innovative TNFα-antagonist. Read More

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Unusual aggregation property of recombinantly expressed cancer-testis antigens in mammalian cells.

J Biochem 2021 Jul 10. Epub 2021 Jul 10.

Department of Interdisciplinary Science and Engineering in Health Systems, Okayama University, Okayama, 700-8530, Japan.

Transient expression of human intracellular proteins in human embryonic kidney (HEK) 293 cells is a reliable system for obtaining soluble proteins with biologically active conformations. Contrary to conventional concepts, we found that recombinantly expressed intracellular cancer-testis antigens (CTAs) showed frequent aggregation in HEK293 cells. Although experimental subcellular localization of recombinant CTAs displayed proper cytosolic or nuclear localization, some proteins showed aggregated particles in the cell. Read More

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Calcium-activated chloride channel is involved in the onset of diarrhea triggered by EGFR tyrosine kinase inhibitor treatment in rats.

Biomed Pharmacother 2021 Jul 8;141:111860. Epub 2021 Jul 8.

Tsumura Kampo Research Laboratories, Tsumura & Co., 3586 Yoshiwara, Ami-machi, Inashiki-gun, Ibaraki 300-1192, Japan. Electronic address:

EGFR tyrosine kinase inhibitors (TKIs) are mainly used to treat non-small cell lung cancer; however, adverse effects such as severe diarrhea represent a major obstacle towards the continuation of EGFR-TKIs therapy. Chloride channels, which control the fluid flow in the intestinal lumen, are proposed as an important target to remediate EGFR-TKIs-induced diarrhea, but the mechanism remains unclear. The aim of this study was to clarify the mechanism underlying EGFR-TKIs-induced diarrhea with a particular focus on the role of intestinal chloride channels. Read More

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Multiplexed functional genomic analysis of 5' untranslated region mutations across the spectrum of prostate cancer.

Nat Commun 2021 07 9;12(1):4217. Epub 2021 Jul 9.

Divisions of Human Biology and Clinical Research, Fred Hutchinson Cancer Research Center, Seattle, WA, USA.

The functional consequences of genetic variants within 5' untranslated regions (UTRs) on a genome-wide scale are poorly understood in disease. Here we develop a high-throughput multi-layer functional genomics method called PLUMAGE (Pooled full-length UTR Multiplex Assay on Gene Expression) to quantify the molecular consequences of somatic 5' UTR mutations in human prostate cancer. We show that 5' UTR mutations can control transcript levels and mRNA translation rates through the creation of DNA binding elements or RNA-based cis-regulatory motifs. Read More

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Improved CRISPR genome editing using small highly active and specific engineered RNA-guided nucleases.

Nat Commun 2021 07 9;12(1):4219. Epub 2021 Jul 9.

Bayer AG, Leverkusen, Germany.

Streptococcus pyogenes (Spy) Cas9 has potential as a component of gene therapeutics for incurable diseases. One of its limitations is its large size, which impedes its formulation and delivery in therapeutic applications. Smaller Cas9s are an alternative, but lack robust activity or specificity and frequently recognize longer PAMs. Read More

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