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We narrowed to 311 results for: cas9 genes

Showing: 271 - 300 of 311 results

  1. Lentiviral Vector Uses and Overview

    Type
    Blog Post
    ...aberrant activation of nearby genes. These safer vectors have become standard in gene therapy. Researchers have...condensed to create the 2nd-generation lentiviral system (Figure 2). The HIV genes that do remain are very...into the genome, they could promote oncogenesis by altering local gene expression. Self-inactivating (SIN... LM, Rebello T. J Gene Med. 2013 Feb;15(2):78-82. PubMed PMID: 23322669. 6. Genetic treatment of a molecular... Jr. Gene Ther. 2010 Feb;17(2):150-7. PubMed PMID: 19847206. Additional Resources on the Addgene Blog ...Genome-Wide Screening Using CRISPR/Cas9 Additional Resources on Addgene.org Check out Our Lentivirus Guide...applications, and they’ve also had success in clinical gene therapy applications. Read on to learn more about...

  2. Tips for arabidopsis transformation

    Type
    Blog Post
    ...Additional resources on the Addgene blog Engineering the plant genome using CRISPR/Cas9 Using pSiM24 for simplifying...transcriptional and translational dynamics of genes and proteins in living plants, to complementing mutant phenotypes...potato contains Agrobacterium T-DNAs with expressed genes: An example of a naturally transgenic food crop....simplifying plant genetic engineering Using arabidopsis in the classroom Resources on Addgene.org Find plant... transformation. There are many motivations to generate transgenic Arabidopsis, from studying transcriptional... floral drip or dip transformation method. The general steps for this method include: Cloning and transforming...Glazebrook, 2002). Find plant plasmid resources at Addgene  Choosing an expression system Choosing an appropriate...

  3. Hot Plasmids - November 2023

    Type
    Blog Post
    ...delivery of genetic cargo  By Alyssa NeuhausTo improve delivery and reduce off-target effects of gene therapy...edits, such as long or highly structured templates. Cas9 variants PE6e–g often had even higher editing efficiencies...system to improve on-target delivery of genetic cargo Optogenetic proximity labeling and TransitID Tracking...continuous evolution and protein engineering to generate seven new prime editors, PE6a–g, that overcome...10.1038/s41467-023-40788-8. PMID 37612276.   Optogenetic proximity labeling and TransitID By Mike Lacy...transcriptional history of a cell. Lin et al. generated a fusion protein called iPAK4 that assembles into...43R] has been one of the hottest antibodies in Addgene’s collection since we began distribution and has...

  4. CRISPR 101: RNA Editing with Cas13

    Type
    Blog Post
    ...the target locus, making them more flexible than Cas9/Cpf1. Some Cas13 enzymes prefer targets with a given...plasmids from this paper at Addgene. Additional Resources on the Addgene Blog Check out our CRISPR Featured...and designated this system REPAIRv1. Using next-generation sequencing, they confirmed A->I editing and found...cytosine deaminase. Three rounds of rational mutagenesis on the ADAR2DD residues that contact the RNA ...RNA-sequencing, the authors performed rational mutagenesis and identified that a S375A mutation in RESCUE...PMID: 29070703. Find plasmids from this paper at Addgene. Abudayyeh, Omar O., et al. “RNA targeting with...PMID: 28976959. Find plasmids from this paper at Addgene. Abudayyeh, Omar O., et al. "C2c2 is a single-...

  5. Split Fluorescent Proteins for Studying Protein-Protein Interactions

    Type
    Blog Post
    ...Split-horseradish peroxidase (Alice Ting Lab 2016)  Split-Cas9 for inducible genome editing (Feng Zhang Lab 2015...resources on the Addgene blog Addgene Blog Biosensors posts Resources on Addgene.org Fluorescent Biosensors...where should you begin? Have no fear, Addgene is here to help!  General Design Figure 1: Two proteins...the FP back together (Figure 1). This method is generally known as Bimolecular Fluorescence Complementation...proteins in Caenorhabditis elegans. Goudeau et al. Genetics. 2021 Cynthia Kenyon mRuby4 mRuby4(1-10), pairs...

  6. The Fluorescent Vegetables in Aptamer Soup

    Type
    Blog Post
    ...cellular RNA to specific intracellular locations using Cas9 and track the localization of the complex. Broccoli...If you’re been following Addgene on #souptwitter or if you just enjoy a warm meal on a winter day, you...molecule into the cell. The fluorescence that is generated allows for qualitative and quantitative measurement...placing the aptamer downstream of a known promoter. Addgene’s Plasmids 101: Aptamer Fluorophores describes the... we’ll take a deeper dive into the content of Addgene’s aptamer soup by exploring the development and ...and applications of fluorescent RNA aptamers in Addgene’s collection. Green fluorescent aptamers Spinach...s group created Spinach2 using site-directed mutagenesis of Spinach. In doing so, they identified the ...

  7. Evolution of Brainbow: Using Cre-lox for Multicolor Labeling of Neurons

    Type
    Blog Post
    ...CRISPR-Cas9 genome editing may be the hot new way to manipulate gene expression, but other gene manipulation...important ways to spatially and temporally control gene expression, especially in in vivo models, and new...at least six generations. In addition to the neuronal pThy1-Brainbow constructs, Addgene also has two ...Brainbow plasmids available from Addgene! Find Brainbow Plasmids @Addgene: Brainbow 1.0, 1.1, 2.1 plasmids...coupling of Cre-lox with FLP-FRT recombination to generate additional recombination events, and the synthesis...and sea anemone mKate2.) They then successfully generated custom antibodies to each of these proteins and...analysis. To achieve even cell labeling, they generated farnesylated derivatives of the fluorescent proteins...

  8. Four Ways to Package Transgenes That Exceed the Size Limit of Adeno-associated Virus

    Type
    Blog Post
    ...approaches increase the size of the gene delivered by splitting the gene into two pieces: part A and part...packaging of large genes in adeno-associated viral vectors results in effective gene delivery in mice. ...how do you fit a big gene into a tiny vector like AAV? By breaking the transgene into smaller pieces. ...acceptor sequence, the 3’ half of the transgene. The full length gene is reconstituted by either the overlapping...Expressing Transgenes That Exceed the Packaging Capacity of Adeno-Associated Virus Capsids. Human Gene Therapy...Chamberlain JS (2011) Gene Therapy of mdx Mice With Large Truncated Dystrophins Generated by Recombination ...retinal gene therapy Learn about the parts of an AAV transfer plasmid Resources on Addgene.org: Check...

  9. Technologies Enabled by NanoLuc® Luciferase

    Type
    Blog Post
    ...enough to detect endogenous tagged genes generated through CRISPR/Cas9 knock-in. Finally, NLuc is very inviting...Fusion Protein, codon-optimized Firefly luciferase genes (e.g., luc2), and NanoLuc® Luciferase are available...such as the dynamics of subcellular structures and gene expression. You must see the movies demonstrating...al. (2016) CRISPaint allows modular base-specific gene tagging using a ligase-4-dependent mechanism. Nature...when employing optogenetic regulators to initiate the event being monitored. Optogenetic regulators respond...wavelengths overlap with optogenetic initiation wavelengths, excitation of the optogenetic regulator leads to... be paired with optogenetic regulators. Many of these tools are available from Addgene and we're excited...

  10. RNA Interference in Plant Biology: New Tools for an Old Favorite

    Type
    Blog Post
    ...Furthermore, gene knockout strategies cannot be used to investigate essential genes. Historically, gene silencing...while gene knockouts have long been considered the “gold-standard” for LOF studies, the high gene copy ...two non-exclusive gene silencing pathways: transcriptional and post-transcriptional gene silencing (PTGS...modifications of complementary gene sequences (Figure 1). As a result, gene silencing can be easily induced...phenotype discrepancies between gene silencing and knockout methods of the same gene. However, recent work has...that is required dsRNA complementary to your gene or genes of interest to trigger RNAi activation. A typical...complementary to your gene target as inverted repeats (Figure 1). With this technique, multiple genes can be simultaneously...

  11. Quick Guide to All Things Lentivirus

    Type
    Blog Post
    ...the env gene containing the rev response element (RRE), an internal promoter, and the desired gene (transfer...-1 genome containing all viral genes with the exception of the env gene (packaging plasmid), and 3) the...used to express your gene of interest either constitutively or inducibly. If your gene of interest is toxic...expression of your gene. The addition of Dox in the culture will then repress your gene of interest. This... Youichi Suzuki (2011). Gene Regulatable Lentiviral Vector System, Viral Gene Therapy, Dr. Ke Xu (Ed.)...harmful genes into a transfer vector. Lentiviral particles expressing proto-oncogenes or oncogenes for instance...safe use in a clinical setting. In this generation, the HIV Tat gene (previously used to drive expression...

  12. How to Deposit Your Plasmids with Addgene

    Type
    Blog Post
    ...Entrez Gene matches to your insert. Select the appropriate gene if it appears in the Entrez Gene list, ...correct Entrez Gene does not appear in the auto-populated list, try entering the gene's Official Full ...known as" aliases in the Gene/Insert box. Once you have selected the appropriate gene from the list, save ...bacterial strain(s) through Addgene. Description – please indicate the gene cloned into your plasmid, any... for the gene or insert. Step 4: Growth and Distribution Please indicate which strain Addgene should propagate... Encodes insert(s) A plasmid with one or more genes, protein coding sequences, or other elements inserted...cloned into it. gRNAs are used for directing CRISPR/Cas9 to a targeted genomic location, while shRNAs are...

  13. Rett Syndrome

    Type
    Collection
    ...and aggressive plan to use gene editing, MECP2 reactivation, RNA editing, gene replacement therapy, RNA ...mutations in the MECP2 gene. Please refer to Addgene's CRISPR Guide for a general introduction to CRISPR...diagnostic criteria have disease-causing mutations in the gene methyl-CpG binding protein 2 ( MECP2 ). MECP2 Rett...caused from loss-of-function mutations in the MECP2 gene. Causative mutations for Rett syndrome typically...global transcriptional regulator of thousands of genes and studies have suggested roles in transcriptional...activity dependent de-repression, chromatin remodeling, gene activation, and more. How loss of MECP2 protein ...suggests the major function of MECP2 is to regulate gene expression either locally or globally. The MECP2...

  14. Fluorescent Proteins 101: Monitoring Cell Mobility Using Fluorescent Proteins

    Type
    Blog Post
    ...The use of other technologies such as optogenetics and CRISPR/Cas9 will help immunologists create even ...molecule (see figure to the right). The excitation generated by these two-photons occurs only at a chosen focal...PubMed PMID: 18431356. Additional Resources on the Addgene Blog Get Tips on Choosing Your Fluorescent Protein...Monitor Protein-Protein Interactions Resources on Addgene.org Check out Our Fluorescent Protein Guide Pages...

  15. Lentiviral Vector Guide

    Type
    Guide
    ...strand of RNA contains packaging genes, regulatory genes, accessory genes, and the long-terminal repeats...shRNA , and overexpressing of oncogenes and other disease-associated genes. Gene Therapy Due to their relative...complex genomes, containing packaging genes and accessory genes specific to each virus type, while retroviruses...require the regulatory genes tat and rev , along with virus-specific accessory genes (for example, vif , ...Packaging plasmid — contains packaging genes gag and pol , and regulatory genes tat and rev ; separated into two... envelope gene, a process called pseudotyping. VSV-G is by far the most common envelope gene, as it has...two plasmids in third generation systems Envelope plasmid — contains packaging gene env (usually VSV-G due...

  16. Antibody Validation for Flow Cytometry

    Type
    Blog Post
    ...endogenous levels (Laflamme et al., 2019). CRISPR-Cas9 approaches are commonly used to produce KO cell ...procedure and application to the major ALS/FTD disease gene C9ORF72. eLife, 8, e48363. https://doi.org/10.7554...101: Validation Resources on addgene.org Addgene's Antibody Guide Addgene's Antibody Protocols... outlined below. Knockout cell line approach Genetic knockout-based validation can conclusively prove...you are working with cells that are amenable to genetic knockout, this represents a very robust approach...to start with a screening procedure in easily genetically modified cell types (top). The specific approach...10.1038/s41598-019-55133-7 More resources from the Addgene blog Introduction to Gating in Flow Cytometry Antibodies...

  17. Fluorescence Titering Assay

    Type
    Protocol
    ...downstream applications. Safety Warnings Lentivirus is generally considered biosafety level 2+. Please ensure that...adherent cells) 0.45 μm polyethersulfone filter, Nalgene, 565-0010 (for viral preps, if prep was not previously...transduced with a range of dilutions of pHAGE-TO-dCas9-3XmCherry . 72 h post transduction, cells were assayed...

  18. Lentivirus Production

    Type
    Protocol
    ...Use this protocol to generate lentivirus...downstream applications such as stable-cell line generation. Workflow Timeline Day 0: Seed 293T packaging...adherent cells) 0.45 μm polyethersulfone filter, Nalgene, 565-0010 Microcentrifuge tubes, Neptune 3745.X...per 10 cm dish). Using transfer plasmid pHAGE TRE dCas9-KRAB (total μg of plasmid DNA 27.8 μg), this would...

  19. Colony Formation Titering Assay

    Type
    Protocol
    ..., BD BD300912 0.45 μm polyethersulfone filter, Nalgene, 565-0010 (for viral preps, if prep was not previously...serial dilutions of the lentiviral vector pHAGE EF1α dCas9-KRAB . The following day, cells were treated with...

  20. Hot Plasmids - August 2020

    Type
    Blog Post
    ...stem cells By Jennifer Tsang CRISPR gene interference (CRISPRi) and gene activation (CRISPRa) are great ways...the plasmids at Addgene! A new tool for synthetic biology uses CRISPRi to control gene expression. This...nucleus and modulate transcription of the mCitrine gene. mCherry is constitutively expressed. Cells are ...selectivity and found that the best motif was from the Lck gene. For marking microtubules of mammalian cells, the... ways to modulate the expression of nearly any gene via specific guide RNA (gRNA) sequences. But these...-IgG1-Fc, Addgene #140700) pMAK463 (NBαMouse-IgK, Addgene #140701) pMAK464 (NBαEGFP, Addgene #140702) ...and the all-in-one dCas9-KRAB and dCas9-VPR targeting plasmids can be found at Addgene. Check out these ...

  21. SARS-CoV-2/COVID-19 Detection Methods Based on CRISPR/Cas

    Type
    Blog Post
    ...amplification kit. In this case, regions of gene S and Orf1ab gene are amplified. RPA reactions also contain...amplification by primers that target the E and the N2 gene of SARS-CoV-2. If the Cas12a-sgRNA complex binds...Approach: FnCas9 Editor Linked Uniform Detection Assay (FELUDA) is a technique that uses FnCas9, a protein...sensitivity of FnCas9, this is also a good way to diagnose single nucleotide variations. Also, FnCas9 cleaves...like CRISPR. Figure 1: An overview of the general schematic of CRISPR/Cas based COVID-19 testing ...protocol for Cas13-based detection of SARS-CoV-2 genetic material (CREST) Sensitivity: 10 copies of RNA ...field-deployable nucleobase detection and identification using FnCas9 (FELUDA) Sensitivity: – 110 femtomolar  Time: Not...

  22. Hot Plasmids: Summer 2024

    Type
    Blog Post
    ...editing, this epigenetic strategy doesn’t induce DNA damage or permanently alter a gene product; it simply...on Prime-editing-Assisted Site-Specific Integrase Gene Editing (PASSIGE), their previously-reported technique...). Efficient site-specific integration of large genes in mammalian cells via continuously evolved recombinases...CRISPRoff system — a programmable epigenetic editor consisting of dCas9 fused to a DNA methyltransferase... is generalizable to a variety of DNA-targeting proteins, so the team created versions using dCas9, zinc...optimizations for prime editing CHARM: A compact epigenetic silencer Recombinant antibodies for marking astrocytes...magnetic beads for scarce macromolecules in heterogeneous samples. bioRxiv 2024.01.21.576499; doi: https...

  23. 8 Great Depositor Research Stories

    Type
    Blog Post
    ...tool for large inserts.  DAP arrays for multiplex gene editing: Guest blogger Qichen Yuan tells us about...Sapozhnikov tells us how to use dCas9 (with no modifications!) as a CRISPR/dCas9-based tool for DNA demethylation...ancestral bias in CRISPR guide library design.  dCas9-directed DNA Demethylation: Guest blogger Daniel... share on our blog, please reach out to news@addgene.org - we'd love to hear from you!       ...

  24. Mobile-CRISPRi: Bringing CRISPRi to Diverse Bacteria

    Type
    Blog Post
    ...antibiotic resistance genes. Genes that are targets of antibiotics are often essential genes which creates a.... ICEBs1 carries the genes needed for conjugation, but can also carry cargo genes, like Mobile-CRISPRi...if that gene is a target of an antibiotic. For example, let’s take a look at the essential gene folA which...targeted essential and non-essential genes with two guides per gene. The library was grown in glucose minimal...essential genes were less represented while bacteria containing guides targeting non-essential genes remained...does not disrupt gene function and is stable and active for greater than 50 generations. Figure ...tools scientists can use to study them. For example, gene knockdown with CRISPR interference (CRISPRi) has...

  25. Bikard Lab - CRISPR Repression Collection

    Type
    Collection
    ...going to bind on the chromosome. When dCas9 binds to a gene, it blocks the progression of the RNA polymerase...Tuning dCas9’s ability to block transcription enables robust, noiseless knockdown of bacterial genes. Molecular...of two genes without cross‐talk. (A) Schematic of the strain expressing two reporters and dCas9 under ...simultaneously. By fusing sfGFP and/or mCherry to genes of interest, it is thus possible to study the effect... protein dCas9 is expressed along a CRISPR array that contains 20 bp "addresses", where dCas9 is going...for quick integration of the aTc-inducible pTet-dCas9 in the attachment site of the phage 186. Detailed...

  26. CRISPR Plasmids - Activate Gene Expression

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    Collection
    ... ID Plasmid Gene/Insert Promoter Selectable Marker PI Publication Bacteria ID Plasmid Gene/Insert Promoter...Drosophila ID Plasmid Gene/Insert Promoter PI Publication Plant ID Plasmid Gene/Insert Promoter Selectable...C. elegans ID Plasmid Gene/Insert Promoter PI Publication Yeast ID Plasmid Gene/Insert Promoter Selectable...transcription of a specific gene. Design your gRNA sequence to direct the dCas9-activator to promoter or ...CRISPR plasmids for activating or upregulating gene expression in various model organisms. CRISPR...or regulatory regions of your gene of interest. If the plasmid that you choose does not also express a ...Transcriptional Regulation Activate Interfere Epigenetics RNA Targeting RNA Targeting RNA Editing Other...

  27. CRISPR Plasmids - Repress Gene Expression

    Type
    Collection
    ... ID Plasmid Gene/Insert Promoter Selectable Marker PI Publication Bacteria ID Plasmid Gene/Insert Promoter...Plant ID Plasmid Gene/Insert Promoter Selectable Marker PI Publication Yeast ID Plasmid Gene/Insert Promoter...dead dCas9, or dCas9 fused to a transcriptional repressor peptide like KRAB, can knock down gene expression...plasmids for transcriptional repression of target genes. CRISPR...with transcription. Design your gRNA to target your gene of interest’s promoter/enhancer or the beginning...Transcriptional Regulation Activate Interfere Epigenetics RNA Targeting RNA Targeting RNA Editing Other...Other Applications Purify Tag Visualize dCas9-FokI Screen Pooled Libraries gRNAs Empty gRNA Vectors gRNAs...

  28. CRISPR Plasmids and Resources

    Type
    Collection
    ...transcription. Epigenetics dCas9 fused to an epigenetic modifier alters chromatin structure/gene expression...Regulation Activate dCas9 fused to an activator peptide activates gene expression. Interfere dCas9 alone, or fused...factors. gRNAs gRNA plasmids to target specific genes. Browse Plasmids by Model Organism Bacteria CRISPR...CRISPR? Experts cover CRISPR topics on Addgene's blog. Addgene's Viral Service : Request ready-to-use viral...Cas enzymes generate double strand breaks (DSBs). Nick Mutated Cas nickase enzymes generate single-strand...CRISPR plasmids available at Addgene. Search by species and CRISPR function. CRISPR...technologies are continously developing and expanding. Addgene works with leading scientists in the field to ensure...

  29. CRISPR Plasmids - Epigenetics

    Type
    Collection
    ...plasmids for epigenetic modification in mammalian or plant systems. Mammalian ID Plasmid Gene/Insert Promoter... target a specific promoter or enhancer for your gene of interest. Available modifications include: histone...Selectable Marker PI Publication Plant ID Plasmid Gene/Insert Promoter Selectable Marker PI Publication...targeted epigenetic modifications, researchers have fused catalytically dead dCas9 to epigenetic modifiers... CRISPR Epigenetics CRISPR Plasmids: Epigenetics Browse CRISPR Plasmids By Function...CRISPR plasmids for targeted epigenetic modification, including histone acetylation/demethylation, and...Transcriptional Regulation Activate Interfere Epigenetics RNA Targeting RNA Targeting RNA Editing Other...

  30. The 12 Days of CRISPR

    Type
    Blog Post
    ...cells🎵 On the 3rd day of CRISPR, Addgene gave to me: dCas9-based gene editing for cleavage-free genomic... of CRISPR, Addgene gave to me: Cas-E-le-ven! (Cas11)🎵 On the 10th day of CRISPR, Addgene gave to me:... day of CRISPR, Addgene gave to me: RADAR from Gao🎵On the 8th day of CRISPR, Addgene gave to me: Functional...CRISPR, Addgene gave to me: updated CRISPR101 blog posts!🎵 On the 4th day of CRISPR, Addgene gave to ...let's get started!  On the 12th day of CRISPR, Addgene gave to me: Noninvasive assessment of gut function...Functional Genetic Variants Revealed by Massively Parallel Precise Genome Editing (CRISPEY!)🎵 On the 7th...7th day of CRISPR, Addgene gave to me: Drag-and-drop genome insertion, without DNA cleavage, with CRISPR-directed...
Showing: 271 - 300 of 311 results