Tag: dCas9

When CASGEVY, the first CRISPR-based therapy, was approved by the FDA in December, it gave CRISPR technology the validation it needed. Any promise it had held before that moment was simply investigational. In this case, the approval of CASGEVY was granted for the treatment of two blood disorders – sickle…

Loss of ASH2L prevents cell proliferation We have studied the molecular and cellular consequences of Ash2l loss in mouse embryo fibroblasts (MEFs) with floxed Ash2l alleles and an inducible Cre-ER recombinase (iMEF-Ash2lfl/fl-Cre-ER). While the knockout of Ash2l was rapid, the downstream effects, including the decrease in promoter-associated H3K4me3, altered gene…

Kalkan, Ali Kerem; Palaz, Fahreddin; Sofija, Semeniuk; Elmousa, Nada; Ledezma, Yuri; Cachat, Elise; Rios-Solis, Leonardo; (2023) Improving recombinant protein production in CHO cells using the CRISPR-Cas system. Biotechnology Advances , 64 , Article 108115. 10.1016/j.biotechadv.2023.108115. Text Improving recombinant protein production in CHO cells using the CRISPR-Cas system.pdf – Accepted Version…

1. Introduction Orientia tsutsugamushi (OT) is an obligate intracellular parasite bacteria and the causative agent of scrub typhus (ST), which is associated with acute febrile illness (AFI) [1] and transmitted by mites through an infected chigger bite (in the larval stage). This disease, which was earlier believed to be endemic…

A feasibility CRISPRi study was performed in post-differentiated iPSC-CMs targeting key genes important in cardiac electrophysiology. Comprehensive analysis using all-optical electrophysiology and a pipeline enabling correlative analysis of functional and molecular data in the same samples helped quantify the CRISPRi gene modulation in this in vitro model. Time-dependent control of…

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…

Rv3907c is the CCA-adding enzyme in Mycobacterium It remains unclear whether the rv3907c gene product, originally annotated as poly(A) polymerase, is in fact the CCA-adding enzyme in Mtb. Rv3907c is composed of three domains, an N-terminal class II polymerase β superfamily domain, a central RNA-binding domain and a C-terminal HD…

Ethics statement All mice procedures in this study were performed under an animal protocol approved by the Institutional Animal Care and Use Committee guidelines of Westlake University. The procedures and protocols for glioma patients were approved by the institutional review board of Beijing Tiantan Hospital. Informed consent was obtained from…

CRISPR-broad framework We developed a procedural pipeline for detecting gRNAs and implemented this in Python as a standalone application (Fig. 1a). For speeding up gRNA selection, we employed multithreading and used big data Python module Pandas. This allowed splitting millions of short sequences for mapping and processing large numbers of uncompressed…

Genome editing technology stands at the forefront of current life science research. However, for its successful translation into clinical applications, the accurate detection of off-target effects is indispensable. Assessing and mitigating off-target effects correctly is a pressing concern. The issue of off-target effects in CRISPR-Cas9 gene editing technology has been…

Statistical methods Statistical tests, test statistics and P values are indicated where appropriate in the main text. Categorical associations were established using the chi-squared test of independence if n > 5 for all categories and Fisherʼs exact test otherwise. For both tests, the Python package scipy.stats v1.5.3 implementation was used64. Multiple hypothesis corrections…

1. Introduction Genome editing has taken a leading position among genome modification technologies in a short time and is now widely used in gene therapy. To date, there are three main systems for genome editing: zinc finger nucleases (ZFNs), transcription activator-like effector nucleases (TALENs), and CRISPR/Cas nucleases. Genome editing has…

Introduction Since the emergence of severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) in 2019, this coronavirus has spread to more than 200 countries, cumulatively infecting 672 million people and causing 6.84 million deaths. Moreover, the emergence of numerous variant strains (α, β, γ, δ, and Omicron) presents a serious challenge for…

Transcriptomics CROP-seq [46] Jurkat Poly-A CRISPRko Cas9 119 – 5798 (+ 1320 cells with NT gRNA) Lentiviral DROP-seq RNA Perturb-seq [43, 44] K562 Barcode CRISPRi dCas9-KRAB UPR epistasis screen: 9 triplet combinations UPR Perturb-seq experiment: 91 Up to 3 gRNAs in a single vector UPR epistasis screen: 15006 UPR Perturb-seq experiment:…

GUS/LUC dual reporter system for detecting CRISPRa efficacy. (a) CRISPRa construct design for OsTRP-like and OsCCR1 genes. (b)Reporter construct design for OsTRP-like and OsCCR1 promoters. (c) CRISPRa efficiency measurement using two reporter genes, GUS and LUC,in Arabidopsis protoplasts. Bar charts represent mean ± sd (n = 3 three independent protoplast…

The Populus genus, commonly known as poplars, cottonwoods, and aspens, consists of approximately 30 tree species native to the northern hemisphere. Because of their diverse usages in landscape, agriculture, bioenergy, and industry, Populus species have been the focus of many tree breeding and genetic improvement programs. Modern biotechnologies, including both genomics and genetic…

  The CRISPR/Cas9 system can be used for knocking out gene expression in vivo or in vitro by using the combination of a sgRNA (single guide RNA) along with Cas9 (dCas9) nuclease. Achieve permanent 100% knockout in your cell line by using Cellecta´s lentivirus-based CRISPR system. Expression of the sgRNA and Cas9…

Cloning of CRISPR/sgRNA lentiviral constructs with fluorescent selection markers A tetracycline-inducible promoter (TRE3G) was used to control the expression of S. aureus dCas9 in a lentiviral vector. To facilitate selection of cells by FACS, pHR:TRE3G-SadCas9-2xKRAB-p2a-tdTomato (Addgene ID #209298) was subcloned from a pHR:TRE3G-SadCas9-2xKRAB-p2a-zeo (A gift from Professor Stanley Qi), where zeocin…

CRISPR-DREAM tool used to activate insufficiently expressed genes and convert skin cells to induced pluripotent stem cells (iPSCs). Published in Nature Methods, Rice University bioengineers developed a tool that activates silent or insufficiently expressed genes using human-derived proteins termed mechanosensitive transcription factors. These naturally enable cells to switch on specific…

Optimization of transfection condition for viral genome staining The transfection was prepared by various concentrations of plasmids expressing DD-dSpyCas9-mCherry-APEX2 (pAPEX2) and sgITR (psgITR) with the transfection agent to stain the rAAV genome inside the cells for TEM. To investigate the target viral genome, a specific staining technique is necessary for…

Design of arabinose-inducible artificial transcription factors A core objective of our work was to develop inducible, heterologous regulators capable of genetically reprogramming gene regulatory networks in Enterobacteriaceae, specifically Salmonella and E. coli. To achieve this, we designed arabinose-inducible ATFs incorporating diverse DBDs originating from CRISPR/dCas9 and plant heterologous TFs (Fig. 1)….

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: PiggyBac dCas9-VPR was a gift from Joanna Wysocka (Addgene plasmid #…

Introduction Cancer is well characterized by genomic instability resulting in structural alterations that build up with tumor progression.1,2 A subpopulation of undifferentiated cancer cells, known as cancer stem cells (CSCs) are found within the tumor bulk. These cells are responsible for cancer initiation, recurrence and show therapeutic resistance and these…

Over the past decade, CRISPR-Cas9 has found widespread application in loss-of-function mutations, but precise genetic engineering for gene correction or gene replacement therapies has lagged behind. In vivo correction using CRISPR-Cas9 to replace genetic mutations by HR is highly challenging, and very few studies have managed to achieve this [31,32]….

Modular iSBH-sgRNA designs enable spatial separation of spacer and trigger-sensing sequences. A. In second-generation iSBH-sgRNAs, RNA triggers are complementary with the iSBH-sgRNA backfolds, thus sgRNA spacers influence RNA trigger sequences. In modular iSBH-sgRNAs, design constrains were eliminated as triggers are only complementary with the iSBH-sgRNA loop and first 15nt of…

Experimental procedures Plasmid design and cloning AAV genome plasmids (Fig. 1a and Extended Data Figs. 1a,g,h and 5a) were based on Addgene plasmid 60231 (ref. 12). To achieve widespread transgene expression, the hSyn promoter was replaced by the ubiquitous CBh promoter (pAS088). For the triple-colour experiments (Extended Data Fig. 1a),…

From RNA interference (RNAi) to CRISPR, there are several methods that researchers can use to manipulate gene function, each with its own strengths and weaknesses. Orthogonal validation – the synergistic use of different methods – makes genetic perturbation studies more robust. Utilizing complementary methods, including RNAi and CRISPR-knockout, -interference and…

Take advantage of Cellecta’s 15+ years of experience making pooled sgRNA Lentiviral Libraries In addition to their ready-to-use, off-the-shelf Genome-Wide CRISPR libraries, Cellecta also provides custom-designed CRISPR sgRNA Pooled Libraries targeting virtually any set of genes and incorporating guide sequences and vector features for a range of screening applications. Custom…

Li, Y., Glass, Z., Huang, M., Chen, Z. Y. & Xu, Q. Ex vivo cell-based CRISPR/Cas9 genome editing for therapeutic applications. Biomaterials 234, 119711 (2020). Article  CAS  PubMed  PubMed Central  Google Scholar  Dong, W. & Kantor, B. Lentiviral vectors for delivery of gene-editing systems based on CRISPR/Cas: current state and…

PPTP-seq development and validation PPTP-seq uses plasmid to integrate each CRISPRi-based TF perturbation and each promoter activity reporter into one construct. Each plasmid contains a CRISPRi cassette that constitutively expresses a single guide RNA (sgRNA) to repress a specific TF in the genome19 and a promoter-reporter cassette to measure the…

Plasmid construction For CRISPR imaging, an sgRNA expression vector, pLH-spsgRNA 2 plasmid (Addgene, Watertown, MA, USA), was amplified, and the oligonucleotides targeting α-satellite DNA repeats (sgRNA1-5)33,49 and 5S rDNA34 were inserted into the BbsI site as previously described16. The sgRNA sequences were as follows: sgRNA1: 5′-CTTTTTGTGGAATTTGCAAG-3′ sgRNA2: 5′-GACTTGAAACACTCTTTTTG-3′ sgRNA3: 5′-TGGAGATTTCAGCCGCTTTG-3′…

Surani MA, Barton SC, Norris ML. Development of reconstituted mouse eggs suggests imprinting of the genome during gametogenesis. Nature. 1984;308:548–50. Article  CAS  PubMed  Google Scholar  McGrath J, Solter D. Completion of mouse embryogenesis requires both the maternal and paternal genomes. Cell. 1984;37:179–83. Article  CAS  PubMed  Google Scholar  Barlow DP, Stoger…

In a study appearing in the Proceedings of the National Academy of Sciences, researchers from Queen Mary University of London investigated the effect of cancer-specific DNA methylation changes on cell physiology. Building off the CRISPR/Cas9 gene editing approach, the investigators developed a method to perform epigenetic editing, which they applied…

In a recent study published in Gene Therapy, researchers utilize the CRISPR-Cas9 system to induce the synthesis of dopamine (DA) in the brains of a rat model for Parkinson’s disease (PD). Study: CRISPR/sgRNA-directed synergistic activation mediator (SAM) as a therapeutic tool for Parkinson´s disease. Image Credit: Sergey Nivens / Shutterstock.com Background The incidence…

The recognition of nucleic acid sequences by proteins is crucial in biology. Protein-DNA binding forms the basis of many biosensing tools, biomolecular engineering techniques, and therapies. To understand and analyze these interactions, it is important to develop simple and sensitive assays that can capture their native behavior while being…

Proteins and nucleic acids interact through specific binding, which plays a critical role in various biological processes. Understanding these interactions is crucial for developing biosensing tools, molecular engineering techniques, and therapies. Scientists have found that nanopores, specifically solid-state nanopores, can be used for resistive pulse sensing to analyze binding events…

DNA-nanostructure barcodes To observe that the CRISPR–dCas9 system is specific enough to detect the single base pair, DNA constructs with different ‘barcodes’ plus an overhang sequence for dCas9 was created. The DNA construct was synthesized from pairing a linearized 7.2 kbp single-stranded (ss) M13mp18 DNA (GuildBiosciences) with 190 complementary oligonucleotides via…

Ndrg2 expression is reduced in tolerogenic DCs To identify potential targets for gene editing in DCs, we compared transcriptomic profiles of treatment induced tolerogenic DCs, which confer a variety of clinical benefits11,12,13,14,15,16,17 with untreated DCs. Bone marrow-derived DCs were cultivated from wild-type (WT) mice (C57/BL6) according to standard protocols25. Vitamin…

WATCH TIME: 4 minutes Welcome to this special edition of Neurology News Network. I’m Marco Meglio. The World Health Organization (WHO) published new editions of the Model Lists of Essential Medicines (EML) and Essential Medicines for Children (EMLc), which now include 3 new therapies for the treatment of multiple sclerosis….

Bourque, G. et al. Ten things you should know about transposable elements. Genome Biol. 19, 199 (2018). Article  CAS  PubMed  PubMed Central  Google Scholar  Chuong, E. B., Elde, N. C. & Feschotte, C. Regulatory activities of transposable elements: from conflicts to benefits. Nat. Rev. Genet. 18, 71–86 (2017). Article  CAS …

In a recent study posted to the preprint server Research Square* while under review for publication in Epigenetics & Chromatin, researchers develop and characterize a highly specific EpiEditing system using catalytically inactivated Cas9 (dCas9) to achieve allele-specific deoxyribonucleic acid (DNA) methylation (ASM). Study: Development of super-specific epigenome editing by targeted allele-specific…

Credit: JUAN GAERTNER/SCIENCE PHOTO LIBRARY/Getty Images CRISPR-based therapies for Alzheimer’s are starting to emerge. At least two possible candidates were reported this week at the Alzheimer’s Association International Conference (AAIC) 2023. One aims to reduce the impact of APOE-e4, the strongest known Alzheimer’s risk gene. The other affects the brain’s…

Ishino Y, Shinagawa H, Makino K, Amemura M, Nakata A. 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(12):5429–33. Article  CAS  PubMed  PubMed Central  Google Scholar  Bhaya D, Davison M, Barrangou R. CRISPR-Cas systems in…

Schematic of odsDNA production and 3 methods of gene knock-in. Credit: Prof. Liang’s group A research group led by Prof. Liang Haojun and researcher Bao Jianqiang from the University of Science and Technology of China (USTC) proposed a novel functional double-stranded DNA (dsDNA) donor that greatly enhances the efficiency of…

Bacterial strains and culture conditions Escherichia coli strain DH5α (Thermo Fisher) was used for the propagation of plasmids, whereas E. coli BL21 DE3 (Novagen) was used for the expression and purification of ResR/McdR protein. Mtb Erdman was obtained from Dr. Ramandeep Singh at THSTI, India, and Mtb H37Rv mc2 790242…

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…

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…

Lesnik E.A., Freier S.M. 1995. Relative thermodynamic stability of DNA, RNA, and DNA:RNA hybrid duplexes: relationship with base composition and structure. Biochemistry. 34, 10807‒10815. Article  CAS  PubMed  Google Scholar  Okou D.T., Steinberg K.M., Middle C., Cutler D.J., Albert T.J., Zwick M.E. 2007. Microarray-based genomic selection for high-throughput resequencing. Nat. Methods….

Amino acid sequence comparisons between GAS and GBS Cas9 identify orthologous active sites Relationships between the amino acid sequence of GBS Cas9 endonuclease and its molecular functions can be deduced from detailed studies of its GAS ortholog, SpyCas9 (Supplementary Fig. 1). As a first step in locating DNA complementary to a…

Phytopathogenic bacterial cytidine base editor system. a A four-module system to make gene specific cytidine base editor. Expression cassettes of guide RNA (gRNA), SacB, and chimeric gene of dead Cas9 (dCas9), cytidine deaminase (CDA1) and DNA uracil glycosylase inhibitor (UGI) each are flanked by the Gateway recombination motifs (attR and…

Professor Liang Haojun, Hefei National Research Center for Physical Sciences at the Microscale, School of Chemistry and Materials Science, University of Science and Technology of China, cooperated with researcher Bao Qiangqiang from the Department of Life Sciences and Medicine, and optimized and proposed the current gene knock-in donors in terms…

Golubnitschaja O, Baban B, Boniolo G, Wang W, Bubnov R, Kapalla M, et al. Medicine in the early twenty-first century: paradigm and anticipation – EPMA position paper 2016. EPMA J. 2016;7:23. Article  PubMed  PubMed Central  Google Scholar  Kropp M, Golubnitschaja O, Mazurakova A, Koklesova L, Sargheini N, Vo T-TKS, et…

Phylogenetic tree construction Tree diagram showing relationships between CDK proteins was constructed from a multi-sequence alignment (MSA) using Geneious95. The “Geneious Aligner”, was used to generate the MSA, and the neighbor joining method was used to construct the tree. All default parameters were used except where otherwise indicated. Combinatorial CRISPR…

doi: 10.1093/nar/gkad355. Online ahead of print. Affiliations Expand Affiliation 1 Institute of Human Genetics, Polish Academy of Sciences, Poznań, Poland. Item in Clipboard Tomasz Woźniak et al. Nucleic Acids Res. 2023. Show details Display options Display options Format AbstractPubMedPMID doi: 10.1093/nar/gkad355. Online ahead of print. Affiliation 1 Institute of Human Genetics,…

1. Introduction Peptides are employed for a multitude of therapeutic approaches and can be obtained from the native organ or tissue in which they are produced as well as through recombinant methods where nucleic acids encoding a candidate gene(s) are delivered to and expressed in cells in vitro. As an…

Ethics statement All experiments in this study comply with guidelines of the U.S. Public Health Service and were approved by the Institutional Biosafety Committee at the University of Pittsburgh. All animal husbandry and experimental procedures were conducted in accordance with U.S. Public Health Service policy and approved by the Institutional…

Plasmids LentiCRISPRv2 (Addgene; 52961) was purchased from Addgene (Cambridge, MA, USA). For HTT gene suppression, LentiCRISPR v2 plasmid with U6 promoter driving sgRNA expression and EF-1 alpha promoter driving the expression of Puromycin-T2A-HA-NLS-dCas9-NLS (or Puromycin-T2A-Flag-NLS-Cas9-NLS) was used. The sgRNA-Cas9 and sgRNA-dCas9 constructs were generated by cloning the CAG repeat-targeting sgRNA…

1 Introduction Abiotic stresses such as drought, heat, cold, salt, pesticides, metals and metalloids deteriorate agro-ecological conditions and hence, negatively impact upon agricultural production. The abiotic stress impairs with regular growth and development of plant plants, thus limiting their yield and quality of produce (Boyer, 1982; Pandey et al., 2017). Plants…

David Allis (1951–2023) was a leading figure in the field of chromatin biology. He inspired many generations of scientists both through his work and his own personal example as a mentor and colleague. His influential ‘histone code’ theory remains an important guiding principle to study and understand gene regulation. In…

Cell culture We cultured mESCs in t2iL medium containing Dulbecco’s modified eagle medium (DMEM, Nacalai Tesque), 2 mM Glutamax (Nacalai Tesque), 1× non-essential amino acids (Nacalai Tesque), 1 mM sodium pyruvate (Nacalai Tesque), 100 U ml−1 penicillin, 100 μg ml−1 streptomycin (P/S) (Nacalai Tesque), 0.1 mM 2-mercaptoethanol (Sigma) and 15% fetal bovine serum (FBS) (Gibco), supplemented with…

Authors: Cholpisit Kiattisewee* (cholpk@uw.edu), Ian D. Faulkner, Jesse G. Zalatan, and James M. Carothers Institutions: University of Washington URLs: Goals CRISPR activation (CRISPRa) and CRISPR interference (CRISPRi) are modular tools that can regulate gene expression of both heterologous and endogenous genes of microorganisms. The project’s goal is to use CRISPRa/i to…

Cell culture Human lung adenocarcinoma cells A549 expressing human ACE2 (A549-ACE2, NR-53821) were acquired from BEI Resources. They were maintained in DMEM/F-12 (1:1, Corning) medium supplemented with 10% FBS (GeneDepot) and blasticidin (100 μM). Normal A549 cells were purchased from ATCC (CCL-185) and cultured in DMEM/F-12 (1:1, Corning) supplemented with 10%…

(A) teloHAEC that express dCas9-VP64 were infected with a lentivirus that carries a sgRNA that targets rs1441755 at the LPL locus. This sgRNA was silent in all our pooled CRISPR screens for all six endothelial phenotypes tested. In the absence or presence of antibiotic selection (Zeocin), LPL_sg08237 does not affect…

NEW YORK – Researchers from Eindhoven University of Technology in the Netherlands have developed an infectious disease molecular testing method that uses bioluminescent proteins they hope to eventually apply at the point of care. The technique, which utilizes recombinase polymerase amplification (RPA) and CRISPR-Cas9 enzymes to detect RNA and DNA…

Researchers from the Wellcome Sanger Institute and collaborators used human stem cells and neurons to investigate what features influence how well CRISPR activation works for different sets of genes. Their study “Massively parallel characterization of CRISPR activator efficacy in human induced pluripotent stem cells and neurons,” which appears in Molecular…

For a paper in Molecular Cell, a team from the Wellcome Sanger Institute and other centers in the UK and China share findings from a barcoded reporter-based analysis of CRISPR activation (CRISPRa) activity in human induced pluripotent stem cells (iPSCs) or differentiated neurons. Starting with iPSCs or neurons containing thousands…

dc.contributor.advisor Chappell, James dc.creator Villegas Kcam, Maria Claudia dc.date.accessioned 2022-12-21T20:09:38Z dc.date.created 2022-12 dc.date.issued 2022-11-08 dc.date.submitted December 2022 dc.identifier.citation Villegas Kcam, Maria Claudia. “Development of trans-acting RNA guided tools to control gene expression in bacteria.” (2022) Diss., Rice University. hdl.handle.net/1911/114170. dc.identifier.uri hdl.handle.net/1911/114170 dc.description.abstract The ability to control gene expression is a…

A gain-of-function screen for regulators of gene silencing We utilized the native Arabidopsis gene FWA as a reporter to screen for regulators of gene silencing. FWA encodes a transcription factor that causes a late flowering phenotype when overexpressed, resulting in a greater number of leaves produced before flowering. In Col-0…

PUBLICATION Authors Weuring, W.J., Dilevska, I., Hoekman, J., van de Vondervoort, J., Koetsier, M., van ‘t Slot, R.H., Braun, K.P.J., Koeleman, B.P.C. ID ZDB-PUB-210819-5 Date 2021 Source The CRISPR journal   4: 575-582 (Journal) Generate reference Registered Authors Koeleman, B. P. C., van ‘t Slot, Ruben, Weuring, Wout Keywords none…

Steve Kanner, Ph.D. has been the Chief Scientific Officer of Caribou Biosciences since 2017, and is responsible for the company’s therapeutic discovery, research, and development activities. Steve received his undergraduate degree in Genetics from the University of California, Berkeley, and his Ph.D. in Immunology and Microbiology from the University of…

In a recent study published in the Microorganisms journal, researchers in China reviewed the uses of bacterial-artificial-chromosome (BAC)-based genome editing techniques in herpesvirus research. Review: Bacterial-Artificial-Chromosome-Based Genome Editing Methods and the Applications in Herpesvirus Research. Image Credit: Panuwach / Shutterstock Herpesviruses are significant human and animal pathogens. Exploring the role of specific…

Lentiviral vector construction and production The lentiviral vector was designed as previously described [13, 41]. LdCV vector contains the following: human codon-optimized S. pyogenes dCas9 which was fused at the C-terminus with VP64-p65-Rta (VPR). LAT-TCS-dCas9-VPR was assembled by fusing LAT (Human cDNA, NM_001014987.2) with dCas9-VPR-Q8 and cloned into a modified…

‘Early life trauma is the strongest risk factor for a range of psychiatric conditions, most particularly depression and anxiety,’ says psychiatrist and neuroscientist Eric Nestler, citing a 2010 paper that found almost all kinds of childhood trauma to be associated with mental illness in adulthood. Interestingly, the analysis suggested that…

Cell culture Normal Human Melanocytes (NHM) were cultured in Dermal Cell Basal Medium (ATCC) with the addition of 5 µg/ml Insulin, 50 µg/ml Ascorbic Acid, 6 mM L-Glutamine, 1.0 µM Epinephrine, 1.5 mM Calcium Chloride, Peptide Growth Factor and M8 Supplement. Dermal fibroblasts (DFs) were isolated from neonatal mice and iPS reprogramming was performed as…

journal contribution posted on 2023-02-14, 19:38 authored by Song Hee Jeong, Hyun Ju Kim, Sang Jun Lee CRISPR interference (CRISPRi) screening has been used for identification of target genes related to specific phenotypes using single-molecular guide RNA (sgRNA) libraries. In CRISPRi screening, the sizes of random sgRNA libraries contained with…

Introduction Lymphoma, a cancer characterized by a malignant tumor of the immune system that originates in the lymph nodes or lymphoid tissue, is one of the most common cancers worldwide. According to GLOBOCAN, the incidence of non-Hodgkin lymphoma (NHL) in both males and females ranked top ten among all cancers…

Zhang H., Qin C., An C., Zheng X., Wen S., Chen W., Liu X., Lv Z., Yang P., Xu W., Gao W., Wu Y. 2021. Application of the CRISPR/Cas9-based gene editing technique in basic research, diagnosis, and therapy of cancer. Mol. Cancer. 20 (1), 126. doi.org/10.1186/s12943-021-01431-6 Article  CAS  Google Scholar …

Sponsored Content by GenScriptNov 16 2022Reviewed by Maria Osipova In a new application for CRISPR/Cas editing tools, researchers at Harvard Medical School, Boston, Massachusetts, have created a CRISPR/Cas9 ribonucleoprotein-based peptide display approach. This technique is hoped to simplify the peptide library analysis workflow by using a new technique called peptide immobilization…

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…

Scientists developed a novel screening methodology for identifying genes that regulate specific microglial functions that may contribute to Alzheimer’s disease (AD) in a recent article published inNature Neuroscience. Characterizing regulatory genes that cause microglia to shift from a healthy state to a diseased state, such as in the brains of…

Arnold, F. H. Design by directed evolution. Acc. Chem. Res. 31, 125–131 (1998). Google Scholar  Packer, M. S. & Liu, D. R. Methods for the directed evolution of proteins. Nat. Rev. Genet. 16, 379–394 (2015). Google Scholar  Drake, J. W., Charlesworth, B., Charlesworth, D. & Crow, J. F. Rates of…

Bartel, D. P. MicroRNAs: Target recognition and regulatory functions. Cell 136, 215–233 (2009). CAS  Article  Google Scholar  Bassett, A. R. et al. Understanding functional miRNA–target interactions in vivo by site-specific genome engineering. Nat. Commun. 5, 4640 (2014). ADS  CAS  Article  Google Scholar  Moore, M. J. et al. miRNA–target chimeras reveal…

doi: 10.15252/emmm.202114797. Online ahead of print. Jessica Giehrl-Schwab #  1   2 , Florian Giesert #  1   2 , Benedict Rauser  1   2 , Chu Lan Lao  3   4 , Sina Hembach  1   2 , Sandrine Lefort  5 , Ignacio L Ibarra  6 , Christina Koupourtidou  3   7 , Malte Daniel Luecken  6 , Dong-Jiunn Jeffery…

This product is a lentiviral plasmid that utilizes the EF1 alpha promoter to drive expression of dCas9-VP64 and blasticidin resistance cassette linked by a 2A peptide (EF1a-dCas9-VP64-2A-Blasticidin) allowing for easy selection following successful transfection or transduction. Use Sigma′s lentiviral dCas9-VP64 plasmid for generation of lentiviral particles and efficient production of…

Date: 11th February 2022 The immune system is a critical biological network of processes that protects an organism from disease, and depends on the ability to distinguish self from non-self, a role driven by antigens.  In humans, T cells respond to antigen stimulation together with the production of cytokines however,…

1. Dunbar, C. E. et al. Gene therapy comes of age. Science 359, eaan4672 (2018). PubMed Google Scholar  2. Wang, D., Tai, P. W. L. & Gao, G. Adeno-associated virus vector as a platform for gene therapy delivery. Nat. Rev. Drug Discov. 18, 358–378 (2019). CAS PubMed PubMed Central Google Scholar  3. Eid, A.,…

Abstract CRISPR-based programmable transcriptional activators (PTAs) are used in plants for rewiring gene networks. Better tuning of their activity in a time and dose-dependent manner should allow precise control of gene expression. Here, we report the optimization of a Copper Inducible system called CI-switch for conditional gene activation in Nicotiana…

CRISPR systems that rely on inactivated Cas enzymes—that is, dead Cas (dCas) enzymes—never looked more alive. They harness the targeting power associated with CRISPR—but not the double-strand cuts. As such, they give researchers new ways to interrogate and manipulate gene function. Possibilities include CRISPR interference (CRISPRi) and CRISPR activation (CRISPRa)…

INTRODUCTION Clustered regularly interspaced short palindromic repeats (CRISPR) are a family of DNA sequences, which constitute a primitive immune system that is responsible for protecting prokaryotic cells from phage infections. New York, Aug. 24, 2021 (GLOBE NEWSWIRE) — Reportlinker.com announces the release of the report “CRISPR Based Therapeutics Market by…

Lead author, Karl Barber with a PICASSO microarray. [Karl Barber, Schmidt Science Fellows] Scientists have used an adaptation of the genome editing technology CRISPR to develop a new peptide display platform that can be used as a tool to identify antibodies in patient blood samples. Researchers from the Howard Hughes…