Tag: GFP

Researchers develop a polyvalent lipid nanoparticle mRNA vaccine against Monkeypox virus

In a recent study posted to the bioRxiv* server, researchers developed a messenger ribonucleic acid (mRNA) technology-based vaccine candidate against the monkeypox virus (MPXV) and evaluated its immunogenicity in animal models. Study: Polyvalent mRNA vaccination elicited potent immune response to monkeypox surface antigens. Image Credit: Wachiwit/Shuterstock Background The mRNA vaccines,…

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Programmable CRISPR-Based Tool ‘Pastes’ in New Genes While Avoiding Double-Strand Breaks

NEW YORK — Researchers at MIT have developed a programmable tool that combines CRISPR and integrases to cut out unwanted stretches of DNA and paste in a replacement. While CRISPR-Cas9 gene editing can make such swaps, it is limited by the need for double-stranded DNA breaks, which can induce undesired…

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Use platform data directly instead of copying

[PATCH] mailbox: mtk-cmdq-mailbox: Use platform data directly instead of copying * [PATCH] mailbox: mtk-cmdq-mailbox: Use platform data directly instead of copying @ 2022-11-02 10:07 AngeloGioacchino Del Regno 0 siblings, 0 replies; only message in thread From: AngeloGioacchino Del Regno @ 2022-11-02 10:07 UTC (permalink / raw) To: jassisinghbrar Cc: matthias.bgg,…

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Nociceptor neurons affect cancer immunosurveillance

Secondary use of biopsies as research specimens The ten melanoma samples used in this study were collected by Sanford Health and classified by a board-certified pathologist. Their secondary use as research specimens (fully de-identified formalin-fixed, paraffin-embedded (FFPE) blocks) was approved under Sanford Health IRB protocol 640 (titled ‘Understanding and improving…

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Affinity microfluidics enables high-throughput protein degradation analysis in cell-free extracts

We designed pDOC to support multiple strategies for analyzing protein degradation in vitro, specifically cell-free systems. The device is based on a MITOMI integrated microfluidic chip, originally developed to quantify protein-ligand interactions at equilibrium24,33. The MITOMI module was modified to contain an array of 16 × 64, 32 × 32…

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Cellular glycan modification by B3GAT1 broadly restricts influenza virus infection

Ethics statement All procedures involving laboratory mice were approved by the Duke University IACUC under the protocol numbers A189-18-08 and A142-21-07. Mice were housed with up to five mice per cage and the ambient room conditions ranged from 70–74 °F and 30–70% humidity with a 12-hour dark/light cycle. Animals were assessed…

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Dual roles of mTORC1-dependent activation of the ubiquitin-proteasome system in muscle proteostasis

Sustained skeletal muscle mTORC1 activity upregulates the ubiquitin proteasome system To understand the molecular mechanisms causing atrophy in muscles with high mTORC1 activation, we first analyzed the transcriptome of the fast-type extensor digitorum longus (EDL) muscle in 3-month-old TSCmKO mice6. A short-term (3 day) treatment with rapamycin was used to…

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Evasion of cGAS and TRIM5 defines pandemic HIV

Cells and reagents HEK293T and U87 cells were maintained in DMEM medium (Gibco) supplemented with 10% fetal bovine serum (FBS, Labtech) and 100 U ml−1 penicillin plus 100 μg ml−1 streptomycin (Pen/Strep; Gibco). THP-1-IFIT1 cells that had been modified to express Gaussia luciferase under the control of the IFIT1 promoter were described previously62. THP-1…

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Inferring and perturbing cell fate regulomes in human brain organoids

Experimental methods Stem cell and organoid culture We used six human iPS cell lines (Hoik1, Wibj2, Kucg2 from the HipSci resource47; 409B2 from the RIKEN BRC cell bank; 01F49i-N-B7 (B7) from Institute of Molecular and Clinical Ophthalmology Basel; and WTC from the Allen Institute) and three human ES cell lines (H1-PAX6YFP…

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Addgene: pLKO5-SgHottip-GFP-CRISPRa

These plasmids were created by your colleagues. Please acknowledge the Principal Investigator, cite the article in which the plasmids were described, and include Addgene in the Materials and Methods of your future publications. For your Materials & Methods section: pLKO5-SgHottip-GFP-CRISPRa was a gift from Suming Huang (Addgene plasmid # 134990…

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Electroporation Protocol Bio Rad Gene Pulser

Unit will be tested before shipping. Rad Gene Pulser different transformations. We autoclave the Freezing Mix and store at room temperature. No limit on the number of engaged members from Commun. It is not necessary to incubate DNA with cells. We have a large inventory of equipment, components and parts…

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Nanobody-based RFP-dependent Cre recombinase for selective anterograde tracing in RFP-expressing transgenic animals

Screening of efficient mCherry-binding protein (MBP) pairs to design Cre recombinase dependent on RFPs First, we aimed to construct Cre recombinase dependent on RFPs based on the reported tool named Cre-DOG30. In this system, N-terminal and C-terminal split Cre recombinase fragments are fused with specific nanobodies for target proteins, and…

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NODULIN HOMEOBOX is required for heterochromatin homeostasis in Arabidopsis

NDX is a heterochromatin-associated factor To understand the genome-wide regulatory roles of NDX, we first analyzed its genomic distribution. For this, we performed chromatin immunoprecipitation sequencing (ChIP-seq) in 10-day old Arabidopsis seedlings expressing N-terminally and C-terminally tagged NDX fusion proteins (flag-NDX/ndx1-1(FRI)/flc-2 and NDX-GFP/ndx1-1(FRI)/flc-2, respectively) expressed from their endogenous promoter21,32. The…

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Live-seq enables temporal transcriptomic recording of single cells

Biological materials RAW264.7, 293T and HeLa cells were obtained from ATCC. RAW264.7 cells with Tnf-mCherry reporter and relA-GFP fusion protein (RAW-G9 clone) were kindly provided by I.D.C. Fraser (National Institutes of Health). The IBA cell line derived from the stromal vascular fraction of interscapular brown adipose tissue of young male…

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Proscillaridin A induces mitochondrial damage and autophagy in pancreatic cancer and reduces the stability of SMAD4 in Panc-1 cells – Hou

Introduction Pancreatic cancer (PC) is one of the most aggressive and lethal cancers (1). Previous studies have shown that aging, environmental and behavioral changes are more closely related to the dramatic increase in the incidence of PC worldwide than genetic factors (2–5). A position paper published by the European Federation…

Continue Reading Proscillaridin A induces mitochondrial damage and autophagy in pancreatic cancer and reduces the stability of SMAD4 in Panc-1 cells – Hou

Transcription-independent regulation of STING activation and innate immune responses by IRF8 in monocytes

Reagents, antibodies, viruses and cells LMW-Poly(I:C), LPS, 2′3′-cGAMP and DMXAA (InvivoGen); hydroxyurea, camptothecin and mitomycin C (MCE); GM-CSF, Flt3L (peproTech); lipofectamine 2000 (Invitrogen); polybrene (Millipore); RNAiso Plus (Takara); HT-DNA (Sigma); SYBR (BIO-RAD); dual-specific luciferase assay kit (Promega); ELISA kit for murine Ifn-β (PBL); ELISA kits for murine IP-10 (Biolegend) were…

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Don’t error if there is no pdata

[PATCH] leds: max8997: Don’t error if there is no pdata * [PATCH] leds: max8997: Don’t error if there is no pdata @ 2022-08-07 10:40 Paul Cercueil 2022-08-08 13:16 ` Andy Shevchenko 0 siblings, 1 reply; 2+ messages in thread From: Paul Cercueil @ 2022-08-07 10:40 UTC (permalink / raw) To:…

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Pooled shRNA Library Screening to Identify Factors that Modulate a Drug Resistance Phenotype

High-throughput RNA interference (RNAi) screening using a pool of lentiviral shRNAs can be a tool to detect therapeutically relevant synthetic lethal targets in malignancies. We provide a pooled shRNA screening approach to investigate the epigenetic effectors in acute myeloid leukemia (AML). The overall goal of the following video is to…

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Researchers use nanotechnology to destroy and prevent relapse of solid tumor cancers

Generation and characterization of DCNV-rAd-Ag. a, Generation of DCNVs derived from adenovirus-infected mature dendritic cells. (1) The genes of tumor-specific antigen were genetically engineered into the adenovirus vector. (2) Recombinant adenovirus infected the immature DC2.4 cells to express the modified antigen on the cell surface and stimulate it. (3) Differentiation,…

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Frontiers | A Mutated Nme1Cas9 Is a Functional Alternative RNase to Both LwaCas13a and RfxCas13d in the Yeast S. cerevisiae

Introduction The clustered regularly interspaced short palindromic repeats (CRISPR)–CRISPR-associated (Cas) protein systems naturally exist in prokaryotes. They are RNA-mediated defense mechanisms to protect bacteria and archaea from invading nucleic acids (Barrangou et al., 2007). CRISPR–Cas systems are classified into two classes (1 and 2). Class 2 CRISPR–Cas require a single…

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fix checkpatch.pl struct should normally be const — DRI Development

Hi Uri, Thank you for the patch! Yet something to improve: [auto build test ERROR on staging/staging-testing] url: github.com/intel-lab-lkp/linux/commits/Uri-Arev/staging-fbtft-fix-checkpatch-pl-struct-should-normally-be-const/20220520-012948 base: git.kernel.org/pub/scm/linux/kernel/git/gregkh/staging.git 4d0cc9e0e53e9946d7b8dc58279c62dfa7a2191b config: arm64-randconfig-r011-20220519 (download.01.org/0day-ci/archive/20220520/202205200821.nJQ0IfFt-lkp@xxxxxxxxx/config) compiler: clang version 15.0.0 (github.com/llvm/llvm-project e00cbbec06c08dc616a0d52a20f678b8fbd4e304) reproduce (this is a W=1 build): wget raw.githubusercontent.com/intel/lkp-tests/master/sbin/make.cross -O ~/bin/make.cross chmod +x ~/bin/make.cross # install arm64 cross compiling tool…

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Tissue distribution and cancer growth inhibition of magnetic lipoplex-delivered type 1 insulin-like growth factor receptor shRNA in nude mice –

The targeted delivery of therapeutic genes into specific tissues, as well as the determination of the biological fate and potential toxicity of nanoparticles, remains a highly relevant challenge for gene-based therapies. Type 1 insulin-like growth factor receptor (IGF-1R), an important oncogene, is frequently over-expressed in lung cancer and mediates cancer…

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Biogenesis, biology and characterization of circular RNAs

Biogenesis, biology and characterization of circular RNAs Kristensen LS et al. The biogenesis, biology and characterization of circular RNAs. Nat Rev Genet. (2019) Summary text Biogenesis and properties of circRNAs Biogenesis of cirRNA Characteristics of circRNAs Discover and analyze circRNAs circRNA genome-wide analysis CircRNA site-specific analysis circRNA visualization Biological functions…

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Study explores SARS-CoV-2 Omicron variant and infected colon epithelial cells

In a recent study posted to the bioRxiv* preprint server, researchers assessed the impact of the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) Omicron variant on infected human cells and organs. Study: SARS-CoV-2 Omicron BA.1 variant infection of human colon epithelial cells. Image Credit: PHOTOCREO Michal Bednarek/Shutterstock Various studies have…

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Plenti Cmv Gfp Recipes

Transplantation of mesenchymal stem cells overexpressing Recipes Details: (Fitchburg, WI). The control plasmid, pLenti–CMV–GFP-2A-Puro and pLenti–CMV–GFP-2A-Puro vectors were purchased from Applied Biological Materials Inc (Richmond, BC, Canada). The in-formation for different antibodies used for this study is provided in Table 1. 2.2 Animals Thirty-nine male, Sprague-Dawley (SD) rats (Charles River,…

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Engineering and validation of a dual luciferase reporter system for quantitative and systematic assessment of regulatory sequences in Chinese hamster ovary cells

Experimental workflow and vector designs One way to validate the all-in-one reporter system is through transient or stable transfection of mammalian cells. Within this framework, the schematics in Fig. 1 illustrate the overall workflow of this study and feature the characteristics of promoters. To summarize, the study involves the engineering and…

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shRNA set against Human ACCSL (NM_001031854.2) (SHH230182)

Description The SureSilencing trade; shRNA Plasmids are designed to specifically knock down the expression of individual genes by RNA interference under either transient (with GFP) or stable transfection (for hygromycin, neomycin or puromycin-resistance) conditions after performance of the appropriate enrichment or selection procedures, respectively. Each vector contains the shRNA under…

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Establishment and Genetic Manipulation of Murine Hepatocyte Organoids

A long-term ex vitro 3D organoid culture system was established from mouse hepatocytes. These organoids can be passaged and genetically manipulated by lentivirus infection of shRNA/ectopic construction, siRNA transfection and CRISPR-Cas9 engineering. The liver is the biggest organ in mammals. It plays very important role in metabolism and detoxification. Although…

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Exploring m6A and m5C Epitranscriptomes upon Viral Infection: an Example with HIV

Hi, in this video we’ll show you how to explore the m6A and m5C epitranscriptomes upon viral infection. To start the experiment, you’ll need 50 million cells for the noninfected and 50 million for the infected condition. In order to obtain a high quality dataset upon analysis, you’ll need to…

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Optogenetic Phase Transition of TDP-43 in Spinal Motor Neurons of Zebrafish Larvae

We describe a protocol to induce phase transition of TAR DNA-binding protein 43 (TDP-43) by light in the spinal motor neurons using zebrafish as a model. So this protocol enables us to induce pathological phase transitions of TDP-43 and address their cellular consequence in the spinal motor neurons in vivo,…

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Connecting science to medicine: tendon-like t

image: (a) Comparison of bio-tendons generated from GFP-iPSC-MSCs and Mkx-iPSC-MSCs. (b) SEM images of the surface layer of GFP-bio-tendon and Mkx-bio-tendon. view more  Credit: Department of Systems BioMedicine, TMDU Researchers from Tokyo Medical and Dental (TMDU) develop artificial tendons in vitro from human stem cells that could fix common tendon injuries…

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Hot Plasmids – February 2022

Every few months we highlight a subset of the new plasmids and viral preps in the repository through our hot plasmids articles. These articles provide brief summaries of recent plasmid deposits and we hope they’ll make it easier for you to find and use the plasmids you need. If you’d ever…

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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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The Microscopy-Based Assay to Study and Analyze the Recycling Endosomes using SNARE Trafficking

Recycling endosomes are part of the endosomal tubular network. Here we present a method to quantify the dynamics of recycling endosomes using GFP-STX13 as an organelle marker. The microscopy-based assay to study and analyze the recycling endosomes using SNARE trafficking. These are skin melanocytes. The black-colored organelles that you see…

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Requirement of Xk and Vps13a for the P2X7-mediated phospholipid scrambling and cell lysis in mouse T cells

Significance The extracellular concentration of adenosine triphosphate (ATP) reaches several hundred micromoles in the inflamed tissues or tumor environment. A high concentration of ATP activates P2X7, a purinergic receptor, and induces the formation of a nonselective cation channel, accompanied by reversible phosphatidylserine (PtdSer) exposure, leading to cell lysis. Here, we…

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Glutamine synthetase licenses APC/C-mediated mitotic progression to drive cell growth

Altman, B. J., Stine, Z. E. & Dang, C. V. From Krebs to clinic: glutamine metabolism to cancer therapy. Nat. Rev. Cancer 16, 619–634 (2016). CAS  Article  Google Scholar  Hensley, C. T., Wasti, A. T. & DeBerardinis, R. J. Glutamine and cancer: cell biology, physiology, and clinical opportunities. J. Clin….

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Generation of NKX2.5 GFP Reporter Human iPSCs and Differentiation Into Functional Cardiac Fibroblasts

doi: 10.3389/fcell.2021.797927. eCollection 2021. Leyre López-Muneta  1 , Javier Linares  1 , Oscar Casis  2 , Laura Martínez-Ibáñez  3 , Arantxa González Miqueo  3   4 , Jaione Bezunartea  5 , Ana Maria Sanchez de la Nava  6   7 , Mónica Gallego  2 , María Eugenia Fernández-Santos  6   7 , Juan Roberto Rodriguez-Madoz  8 , Xabier L Aranguren …

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The Fabrication and Operation of a Continuous Flow, Micro-Electroporation System with Permeabilization Detection

Current therapeutic innovations, such as CAR-T cell therapy, are heavily reliant on viral-mediated gene delivery. Although efficient, this technique is accompanied by high manufacturing costs, which has brought about an interest in using alternative methods for gene delivery. Electroporation is an electro-physical, non-viral approach for the intracellular delivery of genes…

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A cell-based phenotypic library selection and screening approach for the de novo discovery of novel functional chimeric antigen receptors

Cell lines and culture Unless stated otherwise, all tumor target cell lines were sourced from the ATCC via LGC Standards. Human endogenous mesothelin (MSLN)-positive target cell lines H-226 (lung carcinoma, MSLN+, ATCC® CRL-5826™) and AsPC-1 (ATCC CRL-1682), and MSLN-negative Raji control cells (ATCC CCL-86) were cultured in RPMI-1640 Glutamax (Life Technologies,…

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Process Development for the Production and Purification of Adeno-Associated Virus (AAV)2 Vector using Baculovirus-Insect Cell Culture System

In this protocol, AAV2 vector is produced by co-culturing Spodoptera frugiperda (Sf9) insect cells with baculovirus (BV)-AAV2-green fluorescent protein (GFP) or therapeutic gene and BV-AAV2-rep-cap infected Sf9 cells in suspension culture. AAV particles are released from the cells using detergent, clarified, purified by affinity column chromatography, and concentrated by tangential…

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Floxed exon (Flexon): A flexibly positioned stop cassette for recombinase-mediated conditional gene expression

Significance Tools that afford spatiotemporal control of gene expression are crucial for studying genes and processes in multicellular organisms. Stop cassettes consist of exogenous sequences that interrupt gene expression and flanking site-specific recombinase sites to allow for tissue-specific excision and restoration of function by expression of the cognate recombinase. We…

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Identification of a regulatory pathway inhibiting adipogenesis via RSPO2

Integration of APC scRNA-seq data reveals heterogeneity of adipocyte progenitor cells In a previous study9, we defined Lin−Sca1+CD142+ APCs as adipogenesis regulatory (Areg) cells and demonstrated that these cells are both refractory toward adipogenesis and control adipocyte formation of APCs through paracrine signaling. In contrast, Merrick et. al.4 observed that Lin−CD142+ cells…

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Molecular Basis for Retroviral Neuropathogenesis

Murine leukemia-inducing retroviruses (MuLVs) can undergo changes that allow them to cause neurological diseases in rodents, and understanding the molecular basis for these changes will give us insights into the genetic elements required for distinct disease phenotypes. We have been studying PVC-211 MuLV, a variant of the erythroleukemia-inducing Friend MuLV…

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Microtubule Plus-End Dynamics Visualization in Huntington’s Disease Model based on Human Primary Skin Fibroblasts

This protocol is dedicated to the microtubule plus-end visualization by EB3 protein transfection to study their dynamic properties in primary cell culture. The protocol was implemented on human primary skin fibroblasts obtained from Huntington’s disease patients. The Huntington’s disease, HD, is an incurable, neurodegenerative pathology caused by a mutation in…

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Switch to regmap for mmio access

[PATCH] pwm: pwm-mtk-disp: Switch to regmap for mmio access * [PATCH] pwm: pwm-mtk-disp: Switch to regmap for mmio access @ 2022-01-03 15:35 AngeloGioacchino Del Regno 0 siblings, 0 replies; only message in thread From: AngeloGioacchino Del Regno @ 2022-01-03 15:35 UTC (permalink / raw) To: thierry.reding Cc: u.kleine-koenig, lee.jones, matthias.bgg,…

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Sod1 integrates oxygen availability to redox regulate NADPH production and the thiol redoxome

Significance Cu/Zn superoxide dismutase (Sod1) is a key antioxidant enzyme, and its importance is underscored by the fact that its ablation in cell and animal models results in oxidative stress; metabolic defects; and reductions in cell proliferation, viability, and lifespan. Curiously, Sod1 detoxifies superoxide radicals (O2•−) in a manner that…

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Mechanism for DNA invasion of adenoviral Covi

image: Incoming adenovirus particles (AdV) dock at the nuclear pore complexes of a human cell’s nucleus (NPC, dashed line). The cellular enzyme Mind bomb1 (Mib1, grey-white structures) primes them for unlocking and removes protein V (GFP-V, green dots). The uncoated viral DNA genome is then imported into the nucleus. The arrow…

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AAV ShRNA Cloning Service – CD Biospeeds

AAV ShRNA Cloning Service AAV ShRNA Cloning Service Adeno-associated virus (AAV) is a type of parvovirus. Its genome is single-stranded DNA and has the ability to infect both dividing and non-dividing cells. Adenovirus or herpes virus is usually needed to help it replicate and expand in the…

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Single-cell delineation of lineage and genetic identity in the mouse brain

STICR lentiviral library preparation and validation We synthesized a high-complexity lentivirus barcode library that encodes approximately 60–70 million distinct oligonucleotide RNA sequences (STICR barcodes). STICR barcodes comprised three distinct oligonucleotide fragments cloned sequentially into a multicloning site within the 3′ UTR of an enhanced green fluorescent protein (eGFP) transgene under…

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New biosensors allow CRISPR gene edits to be seen with the naked eye

New technology developed by the Department of Energy’s Oak Ridge National Laboratory (ORNL) allows the activity of CRISPR to be seen with the naked eye and an ultraviolet flashlight. In a paper published in ACS Synthetic Biology, scientists describe the development and function of this technology. Their process involves redirecting…

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Watching CRISPR-Cas9 at Work in Real-Time

Detecting the activity of CRISPR gene editing tools in organisms with the naked eye and an ultraviolet flashlight is now possible using technology developed at the Department of Energy’s Oak Ridge National Laboratory. Scientists demonstrated these real-time detection tools in plants and anticipate their use in animals, bacteria and fungi…

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New biosensors shine a light on CRISPR gene editing — ScienceDaily

Detecting the activity of CRISPR gene editing tools in organisms with the naked eye and an ultraviolet flashlight is now possible using technology developed at the Department of Energy’s Oak Ridge National Laboratory. Scientists demonstrated these real-time detection tools in plants and anticipate their use in animals, bacteria and fungi…

Continue Reading New biosensors shine a light on CRISPR gene editing — ScienceDaily

New biosensor sheds light on CRISPR gene editing

ORNL’s biosensor system reveals the CRISPR activity of poplar plants that glow bright green under UV light compared to normal plants that appear red.Credits: Guoliang Yuan / ORNL, US Department of Energy Using technology developed at the Oak Ridge National Laboratory of the Department of Energy, it is now possible…

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New biosensors shine a light on CRISPR gene editing

ORNL’s biosensor system reveals CRISPR activity in poplar plants, which glow bright green under ultraviolet light, compared to normal plants, which appear red. Credit: Guoliang Yuan/ORNL, U.S. Dept. of Energy Detecting the activity of CRISPR gene editing tools in organisms with the naked eye and an ultraviolet flashlight is now…

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Targeted mutagenesis in plants using Beet curly top virus for efficient delivery of CRISPR/Cas12a components

Available online 8 December 2021 doi.org/10.1016/j.nbt.2021.12.002Get rights and content Highlights • An efficient geminiviral vector was developed from Beet curly top virus with a wide host range. • The geminiviral vector delivered CRISPR/Cas components efficiently into plant cells. • A high frequency of both mutagenesis and complete HDR in plant…

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Epistasis shapes the fitness landscape of an allosteric specificity switch

Computational design Protein modeling and design was performed with Rosetta version 3.5 (2015.19.57819)35,37. Python and shell scripts for generating input from Rosetta and analyzing from Rosetta are available at: github.com/raman-lab/biosensor_design The high-resolution TtgR structure co-crystalized with tetracycline was selected as the starting point for computational design (PDB: 2UXH)28. The structure…

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Cristae-dependent quality control of the mitochondrial genome

S. cerevisiae cells distinguish between WT and mutant mtDNA. (A) Schematic illustration of the pedigree analysis. Two Δarg8 yeast strains harboring WT or Δcob::ARG8 mtDNA were mated. Zygotes were isolated, and daughter cells from up to five consecutive generations were separated and placed on free spots on the agar plate…

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MYO10 drives genomic instability and inflammation in cancer

INTRODUCTION Genomic instability often refers to the existence of a variety of DNA alterations, ranging from single nucleotide changes (such as base substitution, deletion, and insertion) to chromosomal rearrangements (e.g., gain or loss of a segment or the whole chromosome) (1). Loss of genome stability can lead to early onset…

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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…

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

High tumor mutation burden and DNA repair gene mutations

Introduction Anaplastic lymphoma kinase (ALK)‑fusion genes represent a small but important part of oncogenic driver mutations in NSCLC, accounting for approximately 3%‑7% of all cases worldwide.1,2 Small molecule tyrosine kinase inhibitors (TKIs) are the standard therapy for ALK-rearranged NSCLC. Crizotinib, a first-generation TKI, is the most widely used targeted drug…

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Disabling de novo DNA methylation in embryonic stem cells allows an illegitimate fate trajectory

The mammalian genome is characterized by widespread methylation of cytosine residues. After fertilization, however, both maternal and paternal genomes undergo extensive demethylation, reaching a low point in the blastocyst (1⇓⇓–4). The embryo genome is then remethylated by the activity of de novo DNA methylation enzymes (5). Mouse embryonic stem (ES)…

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Mini CRISPR Genome Editing System Created

The common analogy for CRISPR gene editing is that it works like molecular scissors, cutting out select sections of DNA. Stanley Qi, assistant professor of bioengineering at Stanford University, likes that analogy, but he thinks it’s time to reimagine CRISPR as a Swiss Army knife. “CRISPR can be as simple as…

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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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Stanford researchers engineer an efficient, multi-purpose, mini CRISPR system

The common analogy for CRISPR gene editing is that it works like molecular scissors, cutting out select sections of DNA. Stanley Qi, assistant professor of bioengineering at Stanford University, likes that analogy, but he thinks it’s time to reimagine CRISPR as a Swiss Army knife. “CRISPR can be as simple…

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Defects in 8-oxo-guanine repair pathway cause high frequency of C > A substitutions in neuroblastoma

Significance The collection of large amounts of whole-genome sequencing data allowed for identification of mutational signatures, which are characteristic combinations of substitutions in the context of neighboring bases. The clinical significance of these mutational signatures is still largely unknown. In neuroblastoma, we showed that high levels of cytosine > adenine…

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ToolGen Files Opposition to CVC Contingent Responsive Preliminary Motion No. 1 | McDonnell Boehnen Hulbert & Berghoff LLP

On June 11th, Junior Party the University of California, Berkeley; the University of Vienna; and Emmanuelle Charpentier (collectively, “CVC”) filed its Responsive Preliminary Motion No. 1 in Interference No. 106,127 to be accorded benefit of priority to U.S. Patent Application No. 13/842,859, filed March 15, 2013, or in the alternative…

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ToolGen Files Opposition to CVC Substantive Preliminary Motion No. 3 to Add Claims in ToolGen Patent | McDonnell Boehnen Hulbert & Berghoff LLP

On May 20th, Junior Party the University of California, Berkeley; the University of Vienna; and Emmanuelle Charpentier (collectively, “CVC”) filed its Substantive Preliminary Motion No. 3 in Interference No. 106,127 (which names ToolGen as Senior Party), asking the Patent Trial and Appeal Board to add claims in ToolGen’s U.S. Patent…

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High-purity production and precise editing of DNA base editing ribonucleoproteins

Abstract Ribonucleoprotein (RNP) complex–mediated base editing is expected to be greatly beneficial because of its reduced off-target effects compared to plasmid- or viral vector–mediated gene editing, especially in therapeutic applications. However, production of recombinant cytosine base editors (CBEs) or adenine base editors (ABEs) with ample yield and high purity in…

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An epigenetic basis of inbreeding depression in maize

INTRODUCTION Charles R. Darwin documented inbreeding depression as growth disadvantages from self-fertilization compared to outcrossing in many plants (1). Prevailing hypotheses suggest that inbreeding depression results from the exposure of deleterious recessive alleles and/or loss of overdominant alleles due to increased homozygosity (2, 3) or reduced recombination frequency in some…

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Can I remove the control in differential expression analysis?

Hi there, Essentially, my experimental design is control vs treatment. Cells were sorted based on fluorescence, so there are 4 different “colors” of treated cells, i.e. red, green, green+red, and blue+green+red. I am interested in how the colors differ from one another. And, I have duplicates for all colors and…

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The LpoA activator is required to stimulate the peptidoglycan polymerase activity of its cognate cell wall synthase PBP1a

Significance Class A penicillin-binding proteins (aPBPs) assemble the bacterial cell wall and are the targets of penicillin and related β-lactam antibiotics. In gram-negative bacteria, the aPBPs require outer membrane lipoproteins to function. However, little is known about how these proteins promote the activity of their cognate synthases in cells. Here,…

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Electroporation ELECTROPORATION It consists of applying short high voltage

ELECTROPORATION It consists of applying short high voltage pulses to a mixture of host cells in suspension and recombinant vectors in solution. These pulses increase the permeability of the membranes, allowing the vectors to pass into the cell. It is a highly efficient method that can be applied to all…

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