Tag: CRISPRa

  The following is an excerpt from an E-book that Twist developed for Genetic Engineering News: The Changing Landscape of CRISPR Screening: A guidebook to the latest CRISPR screening methodologies and technologies. You can download the full E-book here.   Even for seasoned researchers, designing an sgRNA library can be…

Additional Information This liquid Growth Medium is a complete medium optimized to culture select BPS Bioscience cell lines. Optimized for Use With: CRISPRa (SAM) HeLa Cell Line (BPS Bioscience, #78193), CRISPRa (SAM) HepG2 Cell Line (BPS Bioscience, #78194) Article No BPS-78221 Country Availability SE, FI, DK, NO, IS, EE, LV,…

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…

According to a new report published by UnivDatos Markets Insights, the CRISPR gene editing market was valued at more than USD 800 million in 2020 and is expected to grow at a CAGR of around 18% from 2022-2028. The analysis has been segmented into Product Type (CRISPR Products and CRISPR…

About The RoleWe have an exciting opportunity for an experienced bioinformatician who is interested in working at the forefront of the gene editing (CRISPR knock-out, base editing) and gene modulation (CRISPRa, CRISPRi, RNAi) fields. We are seeking a Cambridge-based Senior Bioinformatics Scientist 1 to join our global Bioinformatics team, which…

Integrase-deficient lentivirus (IDLV) enables reverse transcription of an encoded payload from an RNA molecule into a DNA template, which creates a useful yet low-efficiency donor for HDR. We engineered the IDLV genome to include CRISPR ‘cut sites’ that match the intended genomic cut site (Fig. 1). Cas9 ribonucleoprotein electroporated into…

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…

I was wondering if anybody has information on how to choose guides for optimal gene activation/repression using multiplex CRISPR/Cas9s with only two guides? I have tried different combinations e.g. two guides with average activity v.s. one poor guide plus an average guide and got mixed results. Sometimes the latter outperformed…

CLIP enables high-efficiency knock-in into an essential locus To create CLIP, we leverage integrase-deficient lentiviruses (IDLV) to deliver an HDR donor template as a viral RNA genome to cells25,26. IDLV consists of lentiviral components but with a D64V mutation in the integrase, maintaining this enzyme’s ability to package and nuclear…

Background: Epigenetic silencing of tumor suppressor genes (TSGs) is a key feature of oncogenesis in hepatocellular carcinoma (HCC). Liver-targeted delivery of CRISPR-activation (CRISPRa) systems makes it possible to exploit chromatin plasticity, by reprogramming transcriptional dysregulation. Results: Using The Cancer Genome Atlas HCC data, we identify 12 putative TSGs with negative…

A few years ago, a CRISPR activator (CRISPRa) gene-editing technique was successfully developed in a collaborative study carried out at the University of Helsinki, which made it possible to convert adult skin cells to pluripotent stem cells. What makes the CRISPRa technique exceptional is that the process can be implemented…

Responsibilities Location Cambridge, CambridgeshireGB Job ID JR-036129 About The PerkinElmer Life Sciences And Diagnostics Company The PerkinElmer Life Sciences and Diagnostics company – which is to be renamed during the second quarter of 2023 – provides end-to-end solutions that help scientists, researchers and clinicians better diagnose disease and discover new…

Abstract Cas12a CRISPR technology, unlike Cas9, allows for multiplexing guide RNAs from a single transcript, simplifying combinatorial perturbations. While Cas12a has been implemented for multiplexed knockout genetic screens, it has yet to be optimized for CRISPR activation (CRISPRa) screens in human cells. Here we develop a new Cas12a-based transactivation domain…

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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…

Transcriptional dysregulation has been implicated in a number of progressive neurodegenerative diseases. To correct transcriptional misregulation, emerging CRISPR tools such as CRISPR activation (CRISPRa) and CRISPR interference (CRISPRi) could be used. In addition, efficient and precise transcriptional activation in broad brain areas and multiple neuroanatomical structures can be achieved by…

By Eric Rhodes, CEO at ERS Genomics‌ CRISPR/Cas9 genome editing is fast becoming the essential tool for industrial microbial production and chemical synthesis by virtue of its speed, precision, and versatility. CRISPR can drastically improve the efficiency of industrial biotechnology through enzyme engineering and pathway modification and is already being…

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…

Abstract CRISPR-based gene activation (CRISPRa) is a promising therapeutic approach for gene therapy, upregulating gene expression by targeting promoters or enhancers in a tissue/cell-type specific manner. Here, we describe an experimental framework that combines highly multiplexed perturbations with single-cell RNA sequencing (sc-RNA-seq) to identify cell-type-specific, CRISPRa-responsive cis-regulatory elements and the…

Transcribed image text: Which statement describes the results for the transgenic CRISPRa Sim 1+/− mice produced by the TARGATT knockin system? neither promoter nor enhancer activation leads to Sim1 upregulation and reduced weight in transgenic mice only promoter activation leads to Sim 1 upregulation and reduced weight in transgenic mice…

Review . 2023 Mar 23;S1525-0016(23)00145-4. doi: 10.1016/j.ymthe.2023.03.024. Online ahead of print. Affiliations Expand Affiliations 1 Department of Biomedicine, Aarhus University, 8000 Aarhus C., Denmark. 2 Department of Biomedicine, Aarhus University, 8000 Aarhus C., Denmark. Electronic address: bak@biomed.au.dk. Item in Clipboard Review Louise Bendixen et al. Mol Ther. 2023. Show details Display…

(A) Locus view for the CAD locus with nearby gene CNNM2. We provide the position of the sentinel CAD variant (rs11191416) and the putative functional variant identified in the pooled CRISPR screen (rs78260931). The LD proxies and sgRNAs tested are also shown. ATAC-seq and RNA-seq data in resting teloHAEC are…

(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…

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…

New research writes the rules on CRISPR activation  New research starts to write the rulebook on how to effectively use CRISPR activation technology. 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…

Credit: Molecular Cell (2023). DOI: 10.1016/j.molcel.2023.02.011 New research is starting to write the rulebook on how to effectively use CRISPR activation technology. Researchers from the Wellcome Sanger Institute and collaborators have used human stem cells and neurons to investigate the features that influence how well CRISPR activation works for different…

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…

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…

CD Biosciences Drosophila Center focuses on micro-injection, genome editing, cell engineering, genetics, metabolism, neurobiology and other fields. With a mission to provide high-quality and affordable Drosophila products and services, CD BioSciences with a team of experienced experts develops solutions by precise genetic modification of Drosophila as humanized models, as well…

These pooled libraries 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: Mouse Metabolic Gene CRISPRa sgRNA Library was a gift…

In a recent study published in the PLOS Biology, researchers found that leucine-rich repeat-containing protein 15 (LRRC15), a toll-like receptor (TLR)-related cell surface receptor, blocked severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) spike (S) binding. Study: Fibroblast-expressed LRRC15 is a receptor for SARS-CoV-2 spike and controls antiviral and antifibrotic transcriptional…

(A) Schematic of CRISPRa screen used to identify regulators of SARS-CoV-2 spike binding. (B) Ranking of all genes in screen 1 by log2 fold change (LFC) calculated using MAGeCK and plotted using MAGeCKFlute. See also S1 Table. (C) Gene enrichment analysis of screen 1 performed using MAGeCK. Horizontal dotted line…

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…

Abstract Standard transgenic cell line generation requires screening 100-1000s of colonies to isolate correctly edited cells. We describe CRISPRa On-Target Editing Retrieval (CRaTER) which enriches for cells with on-target knock-in of a cDNA-fluorescent reporter transgene by transient activation of the targeted locus followed by flow sorting to recover edited cells….

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…

Abstract The success of CRISPR-mediated gene perturbation studies is highly dependent on the quality of gRNAs, and several tools have been developed to enable optimal gRNA design. However, these tools are not all adaptable to the latest CRISPR modalities or nucleases, nor do they offer comprehensive annotation methods for advanced…

Reviewer #1 (Public Review): In this manuscript, Siepe et al. developed a high-throughput screen designed to identify novel protein-protein interactions in the extracellular human proteome. Their CRISPRa-based method induced the expression of transmembrane receptors such that they could be screened for binding to proteins of interest. Major strengths of this…

Abstract Resistance to endocrine therapy is a common problem in patients with estrogen receptor alpha (ERα) positive breast cancer. In this study, we took a non-biased genome-wide approach to identify novel mechanisms of endocrine resistance using a clustered regularly interspaced short palindromic repeats (CRISPR) activating (CRISPRa) screen. Results from the…

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,…

Scientists at Gladstone Institutes and UC San Francisco (UCSF) say they have co-opted the CRISPR-Cas9 system to forcibly activate genes—rather than edit them—in human immune cells. The method, known as CRISPRa, lets them discover genes that play a role in immune cell biology more thoroughly and rapidly than previously possible,…

SAN FRANCISCO, CA—February 4, 2022—CRISPR genome editing has served as a powerful tool for deleting or altering DNA sequences and studying the resulting effect. Now, researchers at Gladstone Institutes and UC San Francisco (UCSF) have co-opted the CRISPR-Cas9 system to forcibly activate genes—rather than edit them—in human immune cells. The…

Reliable production of high-quality iPS cells enables their increasingly efficient utilization in biomedical applications. Credit: Ras Trokovic, University of Helsinki Researchers at the University of Helsinki have developed a new, faster and more reliable technique for reverting human cells to the stem cell state. Pluripotent stem cells are a key…

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…

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…

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…

Job Summary We have an exciting opportunity in our US office for an experienced bioinformatician who is interested in working at the forefront of the gene editing (CRISPR knock-out, HDR, base editing) and gene modulation (CRISPRa, CRISPRi, RNAi) fields. As a Bioinformatics Scientist 2, you will have the opportunity to…

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)…