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We narrowed to 1,140 results for: Gene

Showing: 221 - 240 of 1140 results

  1. What's New in CRISPR - September 2019

    Type
    Blog Post
    ...either side of the cargo (ex: fluorescent protein gene) on the donor vector. Therefore, targeting requires...developed one of these systems further for simultaneous gene activation and editing. Read the paper in Nucleic... with an NGG PAM motif, averaging 19 targets per gene. The lab has previously used the library with SpdCas9...perform pooled CRISPRi screens to identify essential genes and phage host factors, and identify sequence-specific...Knockout Library is a lentiviral library targeting 26 genes chosen from an analysis of human metastasis cohorts...metastasis CRISPR screen. The Chen Mouse Tumor Suppressor Gene CRISPR Knockout Library is an AAV CRISPR knockout...knockout library targeting mouse tumor suppressor genes (TSG). Find the Mouse Metastasis CRISPR-Cpf1 Double...

  2. Mouse Modeling, Part 2: Breeding and Crossing Mice

    Type
    Blog Post
    ...simple example with the goal is to generate mice that have two modified genes in their genome, and assuming...your targeting vector or gene of interest. 2. Breed your parental mice to generate F1 Once you’ve performed...other will generate approximately 1 in 16 mice that harbor homozygous alleles at both of your genes of interest...your mice. Because their generation involves the integration of a novel gene randomly into their genome...your parental mice were all homozygous for your genes of interest, your F1 mice will be carriers at those... all of your desired genes. Remember Punnett squares from high school biology? They will come in handy...expresses Cre with a strain that expresses a floxed gene of interest. Cre may be expressed globally (in every...

  3. Lambda Red: A Homologous Recombination-based Technique for Genetic Engineering

    Type
    Blog Post
    ...antibiotic resistance genes, as well as non-selectable DNA fragments, such as gene replacements and tags...repressor gene, CI857, is used.  This mutant repressor prevents expression of the recombination genes at low...the red genes needed for recombineering, but also leads to expression of the int and xis genes which are...Substrate DNA design and generation Expression of lambda red recombination genes. Electroporation of substrate... a different genetic background via P1 transduction. Plasmid Expressing lambda red genes from a plasmid...recombinant clones are generated.  A simple way to do this is to express lambda red genes from a plasmid with...using Lambda Red recombineering system to replace a gene of interest with an antibiotic resistance cassette...

  4. PCR Applications for Copy Number Variation Assays

    Type
    Blog Post
    ...number of positive droplets for the gene of interest and reference gene, copy number can be determined through...your experiment?  Complexity of the gene of interest Some gene sequences are harder to amplify and quantify...is a well known fact if a multicopy variant of a gene exists, there must be assays that researchers can... ubiquitously expressed, one-copy ‘housekeeping’ gene), and quantified relative to a 'normal' sample with...to use a reference sample with two copies of the gene (Ma, 2014.) As with all qPCR assays, samples, standard...between 0.80-1.20) would mean you have one copy of the gene; an RQ value of 2 means you have two copies, and... assays, allowing the reference sequence and the gene of interest to be amplified in the same reaction...

  5. Supporting Rare Disease Research with BeHeard 2017

    Type
    Blog Post
    ...disease in which disruption of the smn1 gene results in the degeneration of spinal cord motor neurons. This...Schiavi will use Addgene plasmids to characterize smn1 interactions with other genes. He will use fluorescent...to identify candidate genes as drivers of peritoneal mesothelioma tumorigenesis. They plan to use CRISPR...assess their tumorigenesis. These experiments will serve to validate these candidate genes and allow Nabavi...19,000 live births (1).  While disruption of the smn1 gene is known to cause spinal muscular atrophy, the molecular...fluorescent proteins to monitor the localization of these genes, CRISPR to disrupt their function, and C. elegans... Di Schiavi will be able to better choose which genes and pathways to target in future small molecule ...

  6. Zhang Lab CRISPR Page

    Type
    Collection
    ... information, please visit the Addgene SAM library page . Targeted gene activation using SAM There are...available for mammalian endogenous gene activation using SAM: Addgene plasmids #61422 - #61424 were designed...contain a combination of CRISPR-associated (Cas) genes as well as non-coding RNA elements capable of programming...for the transcriptional activation of endogenous genes. It consists of three components: A nucleolytically...human codon-optimized, and packages both the SaCas9 gene and its single guide RNA (sgRNA) into one plasmid... and LKB1 , the top three significantly mutated genes in lung adenocarcinoma. Delivery of a single AAV...-Cre recombinase and sgRNAs targeting the mouse genes: Kras , p53 , and Lkb1 . This plasmid also contains...

  7. CASTing Off for New Shores in Human Genome Editing

    Type
    Blog Post
    ...and can deliver large sequences and even entire genes. Figure 1: Mechanism of CAST systems. A Cas...evolution using bacteriophages. Researchers moved a gene required for phage propagation to a plasmid that... CAST integration of the promoter could turn the gene on. Once phage success was linked to CAST efficiency...new horizons of evoCAST In the emerging world of gene therapy, highly tailored treatments to specific ...In contrast, a system that can deliver a healthy gene as a single intervention for a variety of loss-of-function...evoCAST is an important step toward that kind of gene treatment. The researchers also compared evoCAST... (Prime-Editing-Assisted Site-Specific Integrase Gene Editing), which combines prime editing with a site-specific...

  8. History of CRISPR Cas - A tale of survival and evolution

    Type
    Blog Post
    ...interspaced palindromic repeats) Cas (CRISPR associated genes) system. This popular genome engineering tool offers... be incorporated into the bacterial genome. Cas genes, which code for polymerases, nucleases, and helicases...adaptive immune system that utilized various cas genes not only to store a record of invading phages but...conserved CRISPR subtypes comprised of more than 40 cas gene families have been characterized. To date, 83% of...Rising to the challenge - Evolution of anti-CRISPR genes in phages The CRISPR/Cas adaptive immune system ...nucleases from re-identifying it (3). Surprisingly, few genes have been identified that neutralize CRISPR/Cas....Earlier this year, expression of 1 of 5 anti-CRISPR genes were found to inactivate the Type I-F CRISPR/Cas...

  9. Expanding the Targeting Scope and Editing Efficiency of Adenine Base Editors

    Type
    Blog Post
    ...plasmid in the host bacterium that expresses gene III. Gene III is under the control of a T7 RNA polymerase... the expression of gene III to create infectious particles. A plasmid-based genetic circuit in the bacteria...activity. A plasmid-based genetic circuit in the bacteria links the expression of gene III to base editor activity... 1: David Liu gives a talk at the Cell Symposia: Gene- and Cell-based Therapies: CRISPR, Stem Cells, and...Cas domains beyond SpCas9. In order to expand the gene-editing applications and targeting scope of ABEs...bacteria links the expression of gene III to base editor activity and editing speed and consists of the... active variant of the base editor, a functional gene III product is produced and can infect bacterial...

  10. Enabling Precision Functional Genomics with the Target Accelerator Plasmid Collection

    Type
    Blog Post
    ...variants in a gene of poorly classified function and a well established cellular signaling gene, thus providing...above to demonstrate how gene expression profiling, systematic tumorigenesis assays, drug epistasis assays... be used as generalizable strategies to map the function of rare cancer variants and genes: High-throughput...bottleneck is that most recurrently mutated cancer genes are rare and most of the individual variants found...well-annotated cellular pathways). It wasn’t clear whether gene-agnostic bioassays could accurately test for molecular...numbers of cancer variants across a large number of genes as systematically as possible. In developing these...assays were designed to complement the one assay per gene approach that has been the historical standard. ...

  11. Viral Vectors 101: Viral Vector Elements

    Type
    Blog Post
    ...them together. Gene expression The reason for most viral experiments is to deliver a genetic cargo, typically...primary plasmid with the delivered cargo (often a gene) is called the transfer plasmid while the other ...component as well. On both sides of this promoter-gene combo are two long terminal repeats (LTRs). These...infiltrate the host cell and deposit viral cargo. Gene expression While AAV often does not integrate into...(more on this below), it’s still used to deliver gene expression constructs. Sandwiched between two inverted... AAV also requires some back up from adenovirus genes – E4, E2a, and VA – which are supplied on a helper...viral replication. More information on these viral genes is available in the adenovirus section below!   ...

  12. Bacterial Toxin-antitoxin Systems as Molecular Biology Tools

    Type
    Blog Post
    ...the ccdB gene is replaced by the gene of interest. As a result, cells that retain the ccdB gene die off...tool. The plasmid pKSCTMF contains the mazF gene and a gene for Zeocin resistance (Figure 2). This MazF-ZeoR...antisense RNA regulates hok gene expression indirectly through the overlapping mok gene." Journal of molecular...associated genes. These systems better ensure that plasmids are passed down through generations (Figure ...application of CcdB is GatewayTM cloning, where the ccdB gene serves as a counterselectable marker in the destination...death. The team used this cassette to knock out genes, knock in a GFP cassette, and introduced site-directed...toxin-antitoxin system to construct a stable synthetic gene circuit that can be used over prolonged periods....

  13. Tips for Using FRET in Your Experiments

    Type
    Blog Post
    ...Express a gene of interest fused to the C-terminus of SCFP3A Amber N1 Yellow Mammalian Express a gene of interest...Express a gene of interest fused to the C-terminus of Amber mVenus N1 Yellow Mammalian Express a gene of interest...your choice of fluorescent fusion protein with your gene of interest: Plasmid Color Expression Description...Aquamarine mCerulean N1 Cyan Mammalian Express a gene of interest fused to the N-terminus of monomeric...Cerulean mCerulean C1 Cyan Mammalian Express a gene of interest fused to the C-terminus of monomeric...monomeric Venus mVenus C1 Yellow Mammalian Express a gene of interest fused to the C-terminus of monomeric...C-terminal His tag SYFP2 Yellow Mammalian Express a gene of interest fused to the C-terminus of SYFP2 Clover...

  14. CRISPR 101: Which Cas9 Do I Choose for My CRISPR Experiment?

    Type
    Blog Post
    ...activators and repressors to target genes. Repressing target genes using dCas9-based repressors Early ...of the gene of interest. dCas9-based repressors are useful when knocking out a particular gene isn’t feasible...repressing target genes in a variety of species and cell types can be found on Addgene’s website. Activating...activating target genes in a variety of species and cell types can be found on Addgene’s website. ...Designing or selecting a sgRNA Addgene may carry a validated sgRNA for your target gene.  Validated gRNAs are ... activate, repress, and even image your favorite gene. With so many Cas9 reagents available, it can be...introducing Cas9-mediated frameshift mutations in the gene of interest. The genomic target for Cas9 and the...

  15. Tips for CRISPR Gene Editing in Mice

    Type
    Blog Post
    ... this gene. Should you cut it at the beginning of the coding sequence to delete the entire gene? Perhaps... in vitro, and using it in mouse embryos to generate gene modified mouse lines. Hopefully this post will...they have been generated (more on that later). Find Validated gRNAs to Target Mouse Genes To begin the ...chances of gene modification in vivo. Find Plasmids for Testing Your gRNAs in vitro at Addgene CRISPR/Cas9...decide to do a gene modification experiment in mice to the day you have your F0 generation (barring life...Haoyi, et al. "One-step generation of mice carrying mutations in multiple genes by CRISPR/Cas-mediated ...blogger Samantha Young. The use of CRISPR/Cas9 for gene editing has expanded since its adaptation for use...

  16. Cancer Research Plasmids and Resources

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    Collection
    ... RAS pathway gene expression vectors for in vitro and in vivo research. TP53 Mutagenesis Library : Pooled... click on a component to find plasmids with your gene of interest. Browse Cancer Research Plasmids by ...of the cell. p53 p53, the most frequently mutated gene in human cancer, is a transcription factor and tumor...Addgene's curated collection of plasmids for cancer research, organized by signaling pathway. Special...Research Cancer Research Plasmids and Resources Addgene has a large variety of resources for use in cancer...

  17. Lentivirus Plasmids

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    Collection
    ...viral particle) and toxicity of the gene product (i.e., if the gene encoded by the transfer plasmid is ... which can be modified to encode a wide range of gene products. The main restrictions on a lentiviral ...chimeric 5’LTR, Any expression cassette (promoter and gene of interest) can be cloned into the plasmid. Nolan...gateway plasmid, constitutive Hygromycin resistance gene coexpression. See article for other versions of ...pULTRA 3rd bi-cistronic expression of EGFP and the gene of interest, also can express shRNA (cloning H1-...mutagensis. The 2nd and 3rd generation lentiviral systems available at Addgene have been designed for increased...used with 2nd and 3rd generation lentivirus technologies. ID Plasmid Generation Type Description PI 12253...

  18. CRISPR 101: Engineering the Plant Genome Using CRISPR/Cas9

    Type
    Blog Post
    ...knockout, activate or repress target genes in plants using the same general experimental design principles ...Resistance by CRISPR/Cas9-Targeted Mutagenesis of the ERF Transcription Factor Gene OsERF922. PLoS ONE 11:e0154027...for targeted modification of known or suspected genes that regulate a desired phenotype. In fact, CRISPR... can also be used to activate or repress target genes in plant cells, respectively. Cas9 expression is...regulate expression of longer RNAs (such as mRNAs for gene expression). Examples of commonly used RNA pol II..., activation and repression of target genes in plants and many of the aforementioned empty gRNA backbones...Agrobacterium tumefaciens as a vehicle to deliver your gene of interest into a target cell line or organism ...

  19. Important Considerations When Using AAVs

    Type
    Blog Post
    ..."Future of rAAV gene therapy: Platform for RNAi, Gene Editing and Beyond." Human Gene Therapy ja (2017...required for the onset of gene expression. However, it has yet to be determined what gene expression looks like...Additional Resources on the Addgene Blog AAV: A Versatile Viral Tool for Gene Expression in Mammals A Match...transport desirable or not desirable? If diffuse gene expression is tolerated, then axonal transport is...Otherwise, using a specific system for controlling gene expression such as Cre-lox, may be sufficient to...tool to use. However, we are starting to see that gene expression really does not take much time at all... normally require second strand synthesis before gene expression begins. ScAAV avoid this rate limiting...

  20. Tips for a 1st Time CRISPR User (by a 1st Time CRISPR User)

    Type
    Blog Post
    ... to make my first CRISPR/Cas9 gene edit. I wanted to inactivate the gene BRAF (a kinase implicated in ...Targeting different regions of the gene is also useful in case the gene has a weird mutation that you didn...each step of performing and analyzing a gene edit using Addgene’s lentiviral CRISPR tools. Schematic...Schematic overview of CRISPR/Cas9 gene editing process. See below for details. The entire process from start...what level of expression was the most efficient for gene editing. A virus encoding GFP plus a gRNA targeting...GFP gRNA, will edit out the virally-delivered GFP gene. So, cells with no Cas9 should appear green and ...multiple gRNAs to target different regions of your gene. I used existing BRAF gRNA viruses, but you can ...
Showing: 221 - 240 of 1140 results