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We narrowed to 311 results for: cas9 genes

Showing: 241 - 270 of 311 results

  1. 10 Great Guest Posts We're Thankful For

    Type
    Blog Post
    ...repression of your target gene, activation and repression of different genes in a single cell, and localization...replication and transcription as well as reporter genes. To learn how you can use minigenomes in your research... them in action. AAV: A Versatile Viral Tool for Gene Expression in Mammals Adeno-associated viruses (... (AAVs) are commonly used to deliver a genes of interest to a variety of cell types in-vivo. These are... editing - all influenced by an understanding of Cas9 biophysics! Casilio: An Adaptive, Multitasking “...localization of a fluorescent signal to your target gene. Read this post to find out more about how the Casilio...are widely used by the optogenetics and chemogenetics communities to activate neurons and neuronal pathways...

  2. Addgene-Seeding Labs Plasmid Grant: Accelerating Science Globally - 5 Scientists, 4 Countries & 3 Continents!

    Type
    Blog Post
    ...in applying gene technology to bacteria and plants. Her research involves cloning cry genes from the soil...plant, Dr. Robertson aims to make use of the CRISPR/Cas9 technology to engineer plants that are able to destroy... shared mission of both Seeding Labs and Addgene. The Addgene-Seeding Labs plasmid grant is a collaborative...of the Addgene-Seeding Labs Plasmid Grant awardees' projects progress. Subscribe to the Addgene blog for.... Additional Resources on the Addgene Blog Learn More About the Addgene-Seeding Labs Plasmid Grant Check...introduce the 5 scientists who were awarded the Addgene-Seeding Labs Plasmid grant this year. These scientists...plant biology, she coordinates the Network for the Genetic Improvement of Cowpea, a knowledge database for...

  3. Hot Plasmids June 2018 - Reverse transcriptase, nanobody, and protein-DNA interaction tools

    Type
    Blog Post
    ...lentiviral plasmids, catalytically inactive Cas9 (dSpyCas9) fused to APEX2 is expressed from a Tet/Shield1...specifically investigated protein binding at the TERT gene, but this method enables the unbiased discovery ...cDNA sequencing and RNA sequencing of complex heterogeneous RNA samples. Furthermore, such high processivity...genomic loci. He made use of catalytically inactive dCas9 by fusing it to an engineered peroxidase, (APEX)...are enriched in the 20 nm radius surrounding the dSpyCas9-APEX2 fusion.  This Sontheimer lab used this technology... technology, which they call C-BERST (dCas9–APEX2 biotinylation at genomic elements by restricted spatial...

  4. Arabidopsis in Education: How the Arabidopsis Biological Resource Center Brings Plant Science to Life

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    Blog Post
    ...Using CRISPR/Cas9 pSiM24: Simplifying Plant Genetic Engineering Resources on Addgene.org Find Plant ...experiments outlined in ABRC’s kits vary by module. As a general guideline, experiments that include growing plants...-12 education.     Additional Resources on the Addgene Blog Learn how BioBuilder can bring Synthetic ...Plant Vectors at Addgene Find Simple Protocols for Introductory Lab Work MoClo Plants Parts Kit ...

  5. Botman-Teusink Yeast FP Collection

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    Collection
    ...constitutive overexpression of FPs and tagging of genes of interest with FPs in yeast. Characterization ... PGK1 promoters and the KanMX marker. ID Plasmid Gene/Insert Selectable Marker Publication Additional ...Yeast Localization and Overexpression (MyLO) CRISPR-Cas9 Toolkit Other Plasmids Bas Teusink Lab Materials... Vickers CE, Bydder SF, Zhou Y, Nielsen LK. Dual gene expression cassette vectors with antibiotic selection...

  6. Rewiring Metabolic Circuitry with CRISPR RNA Scaffolds [Video]

    Type
    Blog Post
    ... delete/overexpress endogenous genes or to introduce heterologous genes, but the trend of research has...) model consists of a catalytically dead Cas9 protein (dCas9) that lacks endonuclease activity coupled...tinkering with regulatory networks and multiplex gene control. However, these approaches are often met...mimic natural transcriptional programs on multiple genes. Modifying the CRISPRi system for multidirectional...programs to differentially alter the levels of five genes that encode biosynthetic enzymes for violacein, ...recruit various transcriptional regulators to alter gene expression at the targeted site. Multiple scaffold...simultaneous, multidirectional and tunable control of gene expression. This strategy can be applied to effectively...

  7. Hot Plasmids and Viral Preps - September 2021

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    Blog Post
    ...systems allow researchers to understand bacterial gene clusters that produce useful natural materials, ...capable of robust expression of high G+C content genes, as validated through monitoring the expression ...end-point sequencing assays. These plasmids use “Cas9-mediated adenosine transient reporter for editing...human mRNA binding proteins with 10 guide RNAs per gene. The lab modified the lentiviral backbone to encode...pooled library from Junjie Chen's lab targets human genes known or suspected to be involved in DNA damage ...Veening is now available with one sgRNA per operon or gene. The sgRNA pool can also target core operons in ...library uses CRISPR cytosine base editors to disrupt gene function by targeting start codons, splice acceptor...

  8. Cultivating Community Science at BosLab

    Type
    Blog Post
    ...journal club discussion on Kevin Esvelt’s CRISPR/Cas9 gene drive paper was very popular. People kept asking... at Addgene.org Order Plasmids from Addgene! Find Free and Open Educational Resources at Addgene! ...BosLab. Once he optimizes plant cell culture and transgenesis he wants to share his knowledge with the community...microbes. We’ve ordered a few yeast plasmids from Addgene and are eager to do our first yeast transformations... @ZebraElement     Additional Resources on the Addgene Blog Experimenting in an Open Source Lab: from...

  9. Fluorescent Protein Travel Awards - FLiPs and Fluorescent Protein Biosensors

    Type
    Blog Post
    ... Intersectin, Cdc42, Raf1, Mek1, Mek2, Cdk5, and Cas9. Xin has demonstrated the utility of these FLiPs...Fatima's work and keep checking back in with the Addgene blog for future award opportunities. Xin Zhou:...when exposed to ultraviolet light. To do so, she generated fusion proteins in which Dronpa domains flanked...Proteins, or FLiPs.  Xin later optimized these first generation FLiPs by mutating Dronpa to lower its aggregation...Chemical Biology! Additional Resources on the Addgene Blog Learn More about Photosactivatable Fluorescent...Fluorescent Protein for Your Experiments Resources on Addgene.org Find Fluorescent Protein Biosensors Browse Plasmids...

  10. Plasmids 101: Inducible Promoters

    Type
    Blog Post
    ... transcription of a target gene, it therefore determines the timing of gene expression and largely defines...activating transcription. Addgene depositors have developed heat shock-inducible Cre and Cas9 for easy genome engineering...synthesis and allowing reporter gene transcription to take place. Addgene also has yellow flame light-regulated...the promoter and ceases repression of the target gene. A simplified lac inducible system is found in many...shock-inducible Hsp70 or Hsp90-derived promoters, in which a gene of choice is only expressed following exposure to...inducible promoters Light is another way to activate gene expression, and two-component systems used in synthetic...promoter pR, preventing expression of a reporter gene. When light is present, YFI is inactive, preventing...

  11. Synthetic Photobiology: Optogenetics for E. coli

    Type
    Blog Post
    ...used with many types of proteins, including Cas9 and dCas9-transcription factors, for which low expression...plasmids from this publication at Addgene. Multichromatic control of gene expression in Escherichia coli...optimized by Tabor’s lab, making light-controlled gene expression in E. coli easier and more robust. Synthetic...used to engineer new pathways with sophisticated gene expression controls. Most cell types do not respond... can be coexpressed for sophisticated control of gene expression. An E. coli light-sensitive two-component... was only about 10-fold, precluding the study of gene expression at very high or very low levels. Schmidl...researchers to better fine-tune the level of output gene expression based on the input light intensity. In...

  12. Improved Plasmid Maps Powered by SnapGene

    Type
    Blog Post
    ...ampicillin resistance gene, you’ll find the full name of the gene, the name of its gene product, and a description...features, such as Cas9, weren’t detected by our in-house software. In fact, SnapGene’s feature library ... plasmid features such as antibiotic resistance genes, selectable markers, promoters, origins, tags, and... Overview of SnapGene powered plasmid maps at Addgene  Three factors make our SnapGene powered plasmid...powered by GSL Biotech's SnapGene Server Software. With the backing of SnapGene’s sequence viewer software...the features you need quickly. SnapGene’s Feature Library - SnapGene’s software seamlessly identifies ...scientists) for suggesting these changes, and the SnapGene and Addgene software development teams for working tirelessly...

  13. Hot Plasmids - December 2019 - CRISPR pooled library, LipoGlo, CRISPRa, and Voltage Imaging Tools

    Type
    Blog Post
    ...from mice that constitutively express Cas9 and screen for genes that affect Th2 differentiation and activation... popular tools for genetic screens, enabling genome-wide interrogation of the genes required for cellular...inserting the NanoLuc luciferase downstream of the apoB gene. Using this reporter, the researchers explore many...-LPs and accelerate the discovery of additional genes and small-molecule probes to aid in the understanding...transduction, now enabling forward genetic screens in cells with few genetic tools. Find the mouse CRISPR ...plasmids at Addgene. Liu et al., Nature Communications, 2019. PubMed PMID: 31451697. Genetically encoded ...responsible for a phenotype can be identified by next-generation sequencing of the genomic DNA. However, some ...

  14. How Addgene Creates Content

    Type
    Blog Post
    ...changes and require updating or retiring. CRISPR/Cas9, for example, has gone through rapid change in the...everything else we do! The goal of Addgene’s educational content Addgene truly believes that sharing speeds...you ever wondered how we choose topics for the Addgene blog? Or why we make certain videos? In this post...You probably won’t be surprised to learn that Addgene’s mission statement extends beyond plasmids, AAVs...topics. What better space to do that than in the Addgene blog?  Fig. 1: One of our favorite content...content collections is our Help Center, help.addgene.org, where you can easily search articles answering ...sure things are clear, accurate, and in line with Addgene style.        Fig. 2: Reference image (...

  15. Luminescent Imaging with Nano-lanterns

    Type
    Blog Post
    ...eNLs as good reporter genes for endogenous protein fusions created using CRISPR/Cas9. eNLs offer high luminescent...plasmids from this paper at Addgene. Additional Resources on the Addgene Blog Learn about Photoactivatable...proteins. To make luminescent imaging a reality, Addgene depositor Takeharu Nagai and colleagues at Osaka...nano-lanterns for use in various applications, including optogenetics, biosensors, and fusion proteins. Solving the...yield, and when transiently expressed in cells, generates much less signal than comparable fluorescent proteins... approach is especially suited to complement optogenetic tools, where light used for excitation can cause...cause unintended activation of the optogenetic system. Biosensors based on Nano-lantern technology are...

  16. AAV Q&A with Tim Miles

    Type
    Blog Post
    ... delivery of CRISPR/Cas9 Tips for a first time AAV user Resources on Addgene.org Find Adenovirus Plasmids...Technique Q1: Is there any way to deliver a long gene (>4.5 kB) by AAV? A: One potential strategy is to...applications in diseases which may be linked to epigenetic changes, such as multiple sclerosis?  A: Basic...an exciting area for future AAV application to epigenetic editing. This area is discussed in our recent...excellent questions.    Additional Resources on the Addgene Blog Using virus in your research - a primer for...

  17. Distribution to Industry

    Type
    Collection
    ...currently available to industry scientists. ID Plasmid Gene/Insert PI Kits Kit name Type PI Description High...Stiewe Use these donor vector libraries for CRISPR/Cas9-mediated saturation genome editing of human TP53...Packaging from Addgene Parkinson's Research from The Michael J Fox Foundation Search Use Addgene's Search to...protection for material providers. Addgene’s deposit agreements allow Addgene to distribute plasmids on behalf...can be submitted to Addgene through your user account or by email to mta@addgene.org . After we review ...A portion of Addgene’s collection is available to qualified for-profit entities. Addgene reserves the ...Addgene has a growing collection of plasmids that are available to industry. Learn more about the available...

  18. Plasmids 101: Screens vs. Selections

    Type
    Blog Post
    ...all known genes. This is a perfect example of an unbiased, reverse screen – all known genes are assayed...of knocking out the genes of interest (or the whole genome), they knock down gene expression temporarily...Sabatini, D.M., Lander, E.S. Genetic screens in human cells using the CRISPR/Cas9 system. Science, 343(6166... a biological process. Players can be proteins, genes, enzymes, drugs, or anything else that plays a role...where there is removal of known factors (such as a genes) and the loss of those factors is assessed. The ...especially useful if you are interested in studying genes whose knockout would be lethal to cells and thus...The two most common methods of manipulation are genetic knockout mutations and treatment with small molecules...

  19. Visualizing Protein Turnover In Situ

    Type
    Blog Post
    ...situ is the knock-in of fluorescent tags into the gene locus of interest. An advantage of this alternative...translation in living single cells. However, before CRISPR-Cas9 these techniques were quite tedious and, even here...studied a synaptic protein that is linked to neurodegeneration. The level of this protein is decreased in...PMCID: PMC4121734. Additional Resources on the Addgene Blog Fluorescent microscopy techniques - Which... tagging strategies Additional Resources on Addgene.org Fluorescent protein plasmids Use FRET to study...protein-protein interactions Deposit plasmids with Addgene! ...

  20. Teaching an Old DOG New Tricks: Controlling Protein Activity with GFP

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    Blog Post
    ...label cells. If GFP could be co-opted to control gene expression, he could then selectively manipulate...used LexA and rTetR systems such that they activate gene expression from different promoters. Moving to an.... GFP expression robustly activated the reporter gene TdTomato, whose expression was absent without electroporated...sites. Placing a lox-STOP-lox cassette upstream of a gene of interest blocks transcription when Cre is absent...present, it removes the STOP cassette, activating gene expression. Although Cre isn’t a modular protein...versions of non-modular proteins like Cre. Could Cas9-DOG be developed soon to allow genome modification...fluorescent proteins as scaffolds for cell-specific gene manipulation.” Cell 154(4) (2013): 928-939. PubMed...

  21. Empty Backbones - Choosing Your Perfect Plasmid Backbone

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    Collection
    ... vector for gene expression pINDUCER21 (ORF-EG) - Inducible lentiviral gene expression ...destination vector for gene expression Lentiviral backbones made by Addgene Addgene has an extensive collection...host cell and are designed for a user to insert a gene of interest. They may contain tags or fusion proteins...These vectors allow you to test the function of your gene of interest in a controlled environment under various...we will assume that you want to express a gene; however, we have included a function section for if you...If you want to drive expression of your favorite gene, you will need a plasmid with a promoter that...mTagBFP2 tag pNIC-GST-TEV-GG - Cloning of target gene with N-terminal GST tag and TEV cleavage Transient...

  22. Overcoming the Challenges of Lentiviral Production

    Type
    Blog Post
    ... containing structural genes and a transfer plasmid carrying the transfer gene. By dividing the viral ...while a single 72h harvest is best for larger genes such as Cas9. See figure 1 for an example of how changing...Talking 'bout my (lentiviral) generation The first step in delivering genes via lentivirus is deciding what... While lentiviral vectors are popular gene delivery tools, producing lentivirus, can pose certain challenges...split the viral genome by removing the viral Rev gene from the packaging plasmid and incorporating it ... the structural elements, envelope, and transfer gene are transfected into a packaging cell line, usually...sufficient titer; this is often the case with large genes. This is due to inefficient packaging of large inserts...

  23. Plasmid-based Recombinant Monoclonal Antibodies: What They Are and Why You Should Be Excited About Them

    Type
    Blog Post
    ...., 2018). Hybridomas can lose gene expression of the mAb-encoding genes or fail to revive after cryopreservation...monoclonal antibodies that are generated in vitro using synthetic genes that are typically expressed from... integrated sequence in a stable cell line. The genes encode the heavy chain and light chain for the antibody...Figure 2: (Left) The light chain and heavy chain genes of the antibody are encoded in a plasmid. (Right...better and better at identifying and cloning the genes for antibody expression. If starting from a protein...sequences of the antibodies and then synthesize the genes that encode those amino acids (Tran et al., 2016...community has created hundreds of useful variants of Cas9. By providing access to the plasmids encoding antibodies...

  24. Adapting Toehold Switches to Detect Zika Virus

    Type
    Blog Post
    ... at Addgene 2. Green, Alexander A., et al. "Toehold switches: de-novo-designed regulators of gene expression...was an in vitro paper-based platform that allowed gene circuits to be deployed out of the laboratory in...combined technologies, we built paper-based synthetic gene networks for the detection of antibiotic resistance...-of-concept work was exciting because it enabled gene circuit-based sensors to be used outside of the ...amplification, new toehold switch designs, CRISPR/Cas9-mediated genotyping, as well as our electronic reader... 3. Pardee, Keith, et al. "Paper-based synthetic gene networks." Cell 159.4 (2014): 940-954. PubMed PMID...Sensor Constructs at Addgene As we report, we have embedded this next generation of toehold switch-based...

  25. Plasmids 101: In-house NGS Plasmid Quality Control Now at Addgene

    Type
    Blog Post
    ...sequence to confirm backbone elements.   Confirming the gene/insert by aligning to NCBI entry or using BLAST....available reference sequence or plasmids containing genes with many modifications. We look for point mutations...amino acid. We also confirm that the species of the gene matches the data associated with the plasmid.   ...Viral Preparations from Viral DNA Extracts. Human Gene Therapy 31:664–678 . https://doi.org/10.1089/hum..., Sanjana NE, Zhang F (2017) Genome-scale CRISPR-Cas9 knockout and transcriptional activation screening...for all incoming plasmids was a big change for Addgene - we had used Sanger sequencing for quality control...control for over ten years. Now, thanks to a generous donation by the Hirsh family, we are able to take...

  26. CRISPR Pooled gRNA Libraries

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    Collection
    ...ID Library Type Species PI Lentiviral Generation gRNAs per gene Total gRNAs Adamson DNA Repair CRISPRi...each containing multiple gRNAs for each target gene. In a CRISPR screening experiment, target cells ...require the use of lentivirus to deliver the gRNA/Cas9 to target cells. Therefore, you must be equipped...are used to knockout, activate or repress target genes. Number of gRNAs - A given CRISPR library may target...target the entire genome or a specific class of genes (subpools). Species - the gRNAs within a given CRISPR...86550 Inhibition Human Weissman 3rd 10 Varies CRISPR/Cas9-assisted Removal of Mitochondrial DNA (CARM) Library...Human Yusa 3rd 5 90,709 Human Interferon-Stimulated Gene CRISPR Knockout Library 125753 Knockout Human Emerman...

  27. Adenoviral Vector Production and Troubleshooting

    Type
    Blog Post
    ...et al., Adenovirus-mediated gene delivery: Potential applications for gene and cell-based therapies in... AdVs are stripped of regulatory genes E1 and E3. Without these genes, AdVs cannot replicate on their ...et al. "Adenovirus-mediated gene delivery: Potential applications for gene and cell-based therapies in...applications and gene therapy: they can be produced at high titers, can accommodate large transgenes, transduce...recombination-competent adenovirus (RCA) can be generated if the E1 gene from the packaging cell line is transferred...for the generation of RCA and reduced immunogenicity. In addition to E1/E3, non-structural genes E2 and...proteins in trans. The Gutless AdV have generated high interest for gene therapy due their increased cloning...

  28. Uncovering Ancestral Bias in CRISPR Research

    Type
    Blog Post
    ...variations in gene expression, they found that for approximately 40% of the identified genes, the genes were not...explanation. Next, they hypothesized that differences in gene expression could explain why some ancestry groups...groups were differentially dependent on certain genes. This didn’t prove to be correct, either. However, ...sequences for sgRNAs that target ancestry-associated genes.   What does the reference genome actually reference... the guide from binding to the genome or prevent Cas9 from cutting, both of which can introduce significant...cases, this may lead researchers to deprioritize genes that could be good targets for therapeutic development...cells’ germline genetic variations. They then wanted to identify cancer-associated genetic dependencies ...

  29. Cloning Mammalian Cells with the Agarose Method

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    Blog Post
    ...overexpress genes of interest; more recently we have also been using Addgene CRISPR vectors to generate cell ...cell lines with knockouts of specific genes. Many years ago, people in the laboratory became frustrated with...cDNA is co-transfected at a favorable ratio to the gene of interest (or, if your protein is fused with a...well plate for further analysis. To assess CRISPR/Cas9 knockout efficacy, we have used both Western blotting...with different levels of expression of our gene of interest. Advantages of the agarose cloning method ...figure 1.   Find Additional Useful Protocols on Addgene's Protocol Pages Agarose cloning method procedure...-well cell plate to a 96-well assay plate. We generally freeze at least 2 ampules each of 4 clones with...

  30. Pooled Library Amplifications

    Type
    Blog Post
    ...   For CRISPR libraries, Genome-scale CRISPR-Cas9 knockout and transcriptional activation screening...Libraries Resources on Addgene.org Addgene's Guide to Plasmid Pooled Libraries  Addgene's Pooled Library Collection...chemical transformations are generally cheaper and easier. One library that Addgene distributes with a chemical...contact us at deposit@addgene.org. Happy amplifying! More resources on the Addgene blog Genome-Wide Screening...If you’re depositing a pooled library with Addgene, you may be surprised to learn that we ask for an ...recombination and loss of plasmid diversity, so Addgene strives to distribute aliquots of the original ...requesting scientist receives a pooled library from Addgene, they’ll need to amplify the sample for further...
Showing: 241 - 270 of 311 results