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We narrowed to 1,140 results for: Gene

Showing: 201 - 220 of 1140 results

  1. Hassle-free 96-well Format Epitope Tagging Using Cas9 Ribonucleoprotein

    Type
    Blog Post
    ... epitope tagging of a large cohort of genes (e.g. tagging genes representing a protein super family). ...knock-in for 96 different genes in a single go (Figure 3). We were able to tag >30 genes/plate in the first ...designed for ~250 genes, we have had about 30% tagging success, although many of these genes were expressed...facilitated easy generation of clonal knock-in lines that we now use for IP and ChIP assays. Tag 96 genes at a time...Efficient CRISPR/Cas9-assisted gene targeting enables rapid and precise genetic manipulation of mammalian .... Additional Resources on the Addgene Blog Plasmids for endogenous gene tagging in human cells Plasmids...Fluorescent tagging of endogenous genes with SapTrap Resources on Addgene.org Browse CRISPR plasmids Check...

  2. 27 Hot Plasmids from 2016

    Type
    Blog Post
    ...an unpublished set of Zika gene plasmids to Addgene’s repository. The genes, from the Zika virus Asian...vectors targeting their genes of interest to generate mAID fusions to endogenous genes. Cell lines with the...targeting a total of 20,121 genes within the human genome (the identity of the genes and target sequences can...expression level for their gene of interest. Although the parts involved in altering gene expression are relatively...contains ~90,000 gRNAs that target ~15,000 genes with 6 gRNAs per gene. The second is a "supplementary" library...library to generate a population of HEK293FT cells expressing a single gRNA targeting a single gene. The gRNA... greatly simplified the process of generating multiple E. coli gene expression plasmids with the Cross-disciplinary...

  3. Adenoviral Delivery of CRISPR/Cas9 Aims to Expand Genome Editing to Primary Cells

    Type
    Blog Post
    ...been used for many years for in vivo delivery and gene therapy and represent one of the most studied viruses...CRISPR-Cas. Figure 1: Schematic of targeted gene knock-out (left) and targeted DNA knock-in (right...binds to target DNA. In the final step of targeted gene knock-out (left), a fragment is deleted from two... This capacity is large enough to carry the Cas9 gene and a gRNA expression cassette in one single viral...Joining (NHEJ) DNA repair mechanism to knock out genes as a result of an insertion, a deletion or a frameshift...with Cas9 and a gRNA designed to target a mutated gene. In a Scientific Reports paper introducing the delivery... CRISPR/Cas9-derived RNA-guided nuclease-induced gene disruption frequencies in the various cell types...

  4. CRISPR Methods for Bacteria: Genome Engineering, CRISPRa, CRISPRi, Base Editing, and More

    Type
    Blog Post
    ...efforts to regulate gene expression were limited to inducible promoters or direct gene knockout. In contrast... of gene expression. They successfully knock down expression of essential and non essential genes in a.... These could be genes, specific polymorphisms, including antibiotic resistance genes. This approach successfully...resistance pCRISPR: carries a spacer targeting the gene of interest and kanamycin resistance E. coli carrying...wild-type sequence persists, Cas9 will cleave the gene of interest, inducing a lethal DSB.     This system.... pCas contains Cas9 and the phage recombination gene lambda Red. pTargetF contains the specific gRNA(... cleavage induces HDR to directly modify a given gene. CRISPR-tranposons By combining CRISPR editing and...

  5. Fluorescent Proteins 101: Luciferase

    Type
    Blog Post
    ...since become a gold standard in gene expression analysis and a luciferase gene (one of many available to choose... Proteins in Nano Lanterns Reporter gene assays A reporter gene, such as luciferase, usually serves as.... The effect of promoters or enhancer regions on gene expression can be determined by detection of the...effect of mutations of these regulatory elements on gene expression. The regulatory element (wild-type or...or mutant) is cloned upstream of the luciferase gene in the plasmid, which is then introduced into the... cells. Expression of luciferase as the reporter gene is measured via its activity to produce light, often...measurement of miRNA efficiency on a target sequence. The gene of interest is cloned into a plasmid downstream ...

  6. A New Optogenetic Tool Based on AraC Controls Gene Expression with Blue Light

    Type
    Blog Post
    ...this plasmid with your gene of interest, you would replace mCherry with your gene of interest. pBLADEONLY...already have a pBAD33-based plasmid expressing your gene of interest, all you have to do is co-transform ...co-transform pBLADEONLY. That way, a previously cloned gene of interest, under the pBAD promoter, can be controlled... Resources on Addgene.org Check out the Molecular Biology Reference Download Addgene's Plasmids 101 eBook...simply more precise. “We show that you have less heterogeneity in the population and you can do all of these...permission.   BLADE is just one of the several optogenetic tools that Di Ventura’s lab and Khammash's lab...lab have created and shared with Addgene. You can find all the plasmids deposited from Di Ventura's lab...

  7. Viral Vectors 101: AAV Variables That Matter

    Type
    Blog Post
    ...Adeno-Associated Viral Vector-Mediated Gene Delivery to the Hypothalamus. Human Gene Therapy, 21(6), 673–682. https...Specificity of gfaABC1D-Driven Transgene Expression in AAV-Mediated Gene Transfer. Molecular Therapy - ...easier to do the validation first.  AAV-mediated gene expression can differ extensively depending on the... cells in far away brain regions expressing your gene of interest. This can cause issues in tracing studies...can occur. Promoters The promoter controls how the gene of interest will be expressed; it can be ubiquitous...chicken-beta-actin or CAG can drive strong early gene expression, but can become methylated over time,...tyrosine-hydroxylase (TH), can limit which cells express your gene. Off-target expression can still occur (Fig. 3) ...

  8. 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... CRISPR Epigenetics CRISPR Plasmids: Epigenetics Browse CRISPR Plasmids By Function...targeted epigenetic modifications, researchers have fused catalytically dead dCas9 to epigenetic modifiers...CRISPR plasmids for targeted epigenetic modification, including histone acetylation/demethylation, and...Transcriptional Regulation Activate Interfere Epigenetics RNA Targeting RNA Targeting RNA Editing Other...

  9. Plasmids 101: Shuttle Vectors

    Type
    Blog Post
    ...while G418 (geneticin) is effective at inhibiting the eukaryotic ribosome. One resistance gene on your vector...used in the lab contain an antibiotic resistance gene to allow for antibiotic selection, but these resistance...aspect of ribosomal function. A puromycin resistance gene on your vector can protect both plasmid-carrying...block protein synthesis, and the neomycin resistance gene protects against any of them. But different aminoglycosides...cause mutations even in cells with the resistance gene, so it isn’t a good fit for all experiments. And...experiment should still include an antibiotic resistance gene for growth in bacteria. Expression You’re almost...systems. But will your cells actually express your gene of interest? Many promoters are species-specific...

  10. Viral Vectors 101: Producing Your rAAV

    Type
    Blog Post
    ...cost-effective manipulation of gene expression in the CNS. Molecular Neurodegeneration, 15(1), Article 1. https...system, in which the required adenoviral helper genes are supplied on two different packaging plasmids...pHelper plasmid containing Ad E2A, E4, and VA RNA genes; and the pRepCap plasmid containing replication ...replication factors and serotype-specific capsid genes.  In this helper-free system, HEK293T cells, expressing Ad...transfected with your transfer plasmid, carrying your gene of interest, and the two helper plasmids, commonly...derived inverted terminal repeat sequences. Human Gene Therapy Methods, 23(1), Article 1. https://doi.org...genomes can limit production of recombinant AAV-2. Gene Therapy, 6(7), Article 7. https://doi.org/10.1038...

  11. Beyond the Blood Brain Barrier: PHP.B Enters the Central Nervous System Via the LY6A Receptor

    Type
    Blog Post
    ...using AAV as a tool for gene expression in mammals. Resources on Addgene.org Search for AAV vectors ...One of the biggest barriers to gene therapy treatment of central nervous system (CNS) diseases is the...has not been identified. While LY6A or other Ly6 gene family members are linked to viral susceptibility...large phenotypic effects such as those that affected gene expression, mRNA splicing, or protein coding regions...as non-permissive to PHP.B CNS transduction. Two genes were associated with the PHP.B permissive phenotype...permissiveness. The Wilson Lab also looked for the gene variant responsible for the PHP.B permissive phenotype...inheritance pattern that suggested there was a single gene that had two codominant versions or alleles that...

  12. Meet the New Addgene Board Members!

    Type
    Blog Post
    ...the Grousbeck Family Chair in Gene Therapy, Director of the Grousbeck Gene Therapy Center, Associate Director...nonprofit companies. His lab focuses on gene therapy work for genetic diseases and over the past few months...deposited AAV constructs for gene delivery. In 2018, he interviewed Addgene co-founder Melina Fan and former...PhD Khandan Baradaran’s expertise in viral safety, gene therapy vector development, vaccine development,...Dimension Therapeutics, she helped build and grow the gene therapy division of the company.  Ultragenyx’s goal... systematically test the function of all 20,000 genes in disease models and then focus drug discovery ...to crises like those and ended up in the field of gene therapy.” He’s also worked on vaccines throughout...

  13. Hot Plasmids and Viral Preps - July 2021

    Type
    Blog Post
    ...for doing gene targeting, reporter generation, controllable gene knockouts, and inducible gene inactivation...Alec Kimmelman targets mouse metabolic genes with ~6 gRNAs per gene.  CRISPR-Act3.0, developed by Yiping... Shirley Liu's lab targets all mouse genes with 10 gRNAs per gene. The guides are optimized to maximize... to develop CRISPR-based approaches for studying gene function in these systems. As a part of this effort...Yiping Qi's lab, allows simultaneous gene activation in plants.  FnCas12a, unlike other Cas12a nucleases identified...opportunities for investigating genetic mechanisms related to morphogenesis, cell fate specification, and...Lab have compiled a new toolkit for performing genetics studies in organoids. The Organoid EasyTag System...

  14. Single-cell tracking of lineage and identity with CellTag

    Type
    Blog Post
    ...scRNA-seq), now allows scientists to identify the gene expression patterns unique to the few cells that...reprogram. Unfortunately scRNA-seq only captures the gene expression of cells at a single time point, making...at the single-cell level, thus linking ancestral gene expression patterns with reprogramming outcomes ...constructing lineage trees and comparing them with the gene expression changes of more than 100,000 cells undergoing...partially reprogrammed and re-expressed fibroblast genes. Most cells ended up taking the ‘dead-end’ path....cells with shared CellTagD13 tags often had similar gene expression pattern changes and reprogramming outcomes...tracing experiment, the Morris lab identified a novel gene, Mettl7a1, whose expression helped coax cells towards...

  15. CRISPR-based Genome Editing Systems For Editing in E. coli

    Type
    Blog Post
    ...multiple terminators can serve to interrupt genes (or gene clusters) in metabolic engineering. So far,...pTargetF with the ccdB gene containing BsaI restriction enzyme sites at its ends to generate pEcgRNA. The ccdB...to successfully delete genes of interest in BL21(DE3) using the newly generated pEcCas/pEcgRNA system....used MUCICAT to optimize the gene dosage of glucose dehydrogenase and generated a strain that could achieve...self-curing, an arabinose inducible lambda-red gene, and an IPTG-inducible sgRNA sequence targeting the...non-temperature-sensitive replicon, and 3) added the sacB gene for counterselection to yield pEcCas. In pTargetF... of its E. coli host, and the addition of a sacB gene renders it beneficial for plasmid clearance. It ...

  16. Viral Vectors 101: Integrase-Deficient Lentiviral Vectors

    Type
    Blog Post
    ...Gag, Pol, Rev, and Tat genes. Specifically for our knowledge here, the Pol gene encodes three separate...Insertional mutagenesis combined with acquired somatic mutations causes leukemogenesis following gene therapy...insertional mutagenesis. This integration could lead to an undesirable interruption of a critical gene. Because...Insertional mutagenesis is especially concerning in clinical settings, as evidenced by retroviral gene therapy...transcriptase), and IN (integrase). These viral packaging genes are required for the production of lentiviral particles...portion does not contain any of the viral packaging genes, the natural cycle of the lentiviral vector ends...transcriptionally active, leading to transient expression of gene products (Wu et al., 2004). Over time, the expression...

  17. Using CRISPR/Cas9 to Edit Disease Out of the Genome

    Type
    Blog Post
    ...2020. . “Advances in CRISPR/Cas-based Gene Therapy in Human Genetic Diseases.” Theranostics. https://doi.org...treat AD using CRISPR/Cas9. They are targeting the genes responsible for both early onset and late onset ...Sciences found that mice with a dominant mutation in a gene that causes cataracts could be rescued by coinjection...dominant cataract disorder caused by a single copy of a gene known as Crygc. When all was said and done, the ...able to demonstrate functional correction of the gene in clonally expanded organoids. “CRISPR in adult...fibrosis may not be the most likely candidate for gene therapy, however, because the disease affects multiple...proof-of-principle for use of CRISPR to correct single-gene conditions. Remaining hurdles in CRISPR biology...

  18. University of Florida Serotype Testing Panel for the Eye and Brain

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    Collection
    ...vectors for human gene therapy of glaucoma and their reversed cell entry characteristics. Gene Ther . 2020 ...cells by a mutant adeno-associated viral vector. Hum Gene Ther Methods . Feb;25(1):72-82. PMID: 24191859 Ye...Adeno-Associated Virus Vector Expressing Retinoschisin. Hum Gene Ther Clin Dev . Sep;26(3):165-76. PMID: 26390090... Display Exceptional Characteristics for Retinal Gene Therapy. Mol Ther . Jun 3;28(6):1464-1478. PMID:...: Viral Vectors As part of our Viral Service , Addgene is producing ready-to-use viral preparations from...preparations were produced with the pTR-UF11 plasmid ( Addgene #157970 ). Viral preparations are formulated in...

  19. Addgene’s GIPHY Page is Here to Help You Express Yourself (and Your Gene of Interest)

    Type
    Blog Post
    ... on a break from expressing some genes of interest).   Blugene is so excited they can’t stop dancing...GIFs, feel free to let us know on Twitter @addgene! Addgene is always looking for new ways to help you...We’re making some moves over here at Addgene...and they’re looping? What’s the deal with looping...coworkers, labmates, and friends as well!   Addgene is sharing so much more than plasmids these days...GIPHY   You can see for yourself at giphy.com/addgene. But that’s not all! You’ll be able to use these...Twitter, Instagram, TikTok and more. Just type “addgene” into the GIF search bar to find that perfect response....   This could be you after watching Addgene’s protocol videos. via GIPHY   If you’re also...

  20. FLEx Technology and Optogenetics: Flipping the switch on gene expression with high spatial and temporal resolution

    Type
    Blog Post
    ...reporter gene (e.g., mCherry) to easily detect opsin-expressing cells. The resulting fusion gene is inserted...downside in optogenetics (Belzung et al., 2014). This could be overcome by fusing opsin genes to cell-type...Krook-Magnuson, 2018). Optogenetic FLEx vectors contain a strong promoter upstream of an opsin gene (e.g., channelrhodopsin...2014). Microbial rhodopsins are encoded by opsin genes, and include bacteriorhodopsins, halorhodopsins,...light-sensitive, so the selective expression of opsin genes in targeted cells makes it possible to control neuronal...viruses (rAAVs) have become increasingly popular for gene delivery in the central nervous system due to their...those usually drive weak expression of downstream genes. Strong cell-specific promoters, on the other side...
Showing: 201 - 220 of 1140 results