Skip to main content
Addgene

Enter a sequence to perform a BLAST-based search of the Addgene plasmid sequence database.

Information

In Category

We narrowed to 920 results for: Gene

Showing: 81 - 120 of 920 results

  1. Adenoviral Vector Production and Troubleshooting

    Type
    Blog Post
    Published
    June 14, 2018, 11:44 a.m.
    ...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...

  2. Quick Guide to Working with Drosophila Part 1: Getting Started with Flies

    Type
    Blog Post
    Published
    July 13, 2017, 2:30 p.m.
    ... you started. Any given gene’s webpage will have information on where the gene is expressed within the...from mutations in the gene, and whether any publications have mentioned the gene. The Bloomington Drosophila...also has collections of flies for in vivo gene knock down and gene expression. In the next post, you’ll ...favorite gene (YFG) in Drosophila (and even if there’s not!) the wealth of Drosophila genetic tools available...PhD student at Harvard University. Do you have a gene of interest but have run into a wall trying to study...it? It happens. Is it an evolutionarily conserved gene? Can you find an ortholog in the Drosophila genome...a chance of seeing a phenotype since additional genes that can do the same function are less likely to...

  3. CRISPR 101: Targeting Non-Coding RNAs with CRISPR/Cas9

    Type
    Blog Post
    Published
    Nov. 14, 2024, 2:15 p.m.
    ...nearby genes and its role in gene expression, though this may still unexpectedly affect nearby genes (Figure...protein-coding genes thanks to systems like CRISPR/Cas9 that allow them to manipulate full genes and observe..., or permanently disable, protein-coding genes by generating insertions or deletions in the DNA sequence...only 2% of our genome is made of protein-coding genes, and you might be wondering what the rest of our...manipulate and study in comparison to protein-coding genes (Rinn and Chang 2012). Scientists have determined...multiple gRNAs poses the risk of disrupting adjacent genes, especially those regulated by or overlapping with...). The interactions between ncRNAs and adjacent genes are complex and not yet fully understood, so it ...

  4. 28 Hot Plasmid Technologies from 2015

    Type
    Blog Post
    Published
    Dec. 23, 2015, 3:30 p.m.
    ... cassette to the gene of interest. The gene to be expressed is amplified using a gene-specific forward...sgRNA-targeted gene, resulting in increased activation of endogenous transcription of the target gene. The brilliant...of downstream genes. Nihongaki et al describe the targeted activation of endogenous genes and detail the...efficiency and gene expression. The pUCXKT vector ensures that all screened plasmids contain a gene variant*...resistance gene (missing the first two codons). Two stop codons are inserted between the truncated gene and ... inducible, gene knockdown with shRNA or shRNA-miR30, (2) tamoxifen-inducible CreERT2 gene deletion, (...on target gene activation. The novel dCas9-VPR activator is capable of enhancing target gene expression...

  5. CRISPR 101: Drug discovery and CRISPR

    Type
    Blog Post
    Published
    May 4, 2023, 1 p.m.
    ...several candidate inhibitors of gene X in a screen. You generate a gene X knockout in a cancerous background...inhibit, activate, or knockout many genes at a time to identify ‘hits’ - genes that affect disease progression...some of the candidate genes! Disease model generation CRISPR has made the generation of genome edits accessible.... These hits can reveal individual genes or sometimes even genetic pathways that are worth investigating...known – repeat expansion, gene knock out, etc. – CRISPR can be used to generate cell line or animal models...approaches. The first route involves identifying a gene/pathway to develop a drug for (aka a druggable target...straightforward causes (a specific mutation in a known gene = disease onset) may be best served by this type...

  6. Synthetic promoter AAVs for cell-type specific expression in retinal cells

    Type
    Blog Post
    Published
    March 12, 2020, 1:15 p.m.
    ... popular tool for gene delivery, but it has a drawback: how do you ensure your gene goes where you want...cell-type specific genes from mice. Strategy #2 (ProB): Create promoters using phylogenetically conserved sequences...projections. Generating animal models: Transgenic animals allow for the expression of genes in particular...Additional resources on the Addgene blog Read more about using AAV for retinal gene therapy Find more about...want it to? Knowing that a gene is expressed in a particular cell type is important not only for translational...translational research, such as gene therapy, but also basic research. To improve cell-type specificity of... transcription initiation sites of at least two genes with high expression and high cell-type specificity...

  7. Plasmids 101: Walkthrough of Addgene’s Snapgene-Powered Quality Control Process

    Type
    Blog Post
    Published
    Feb. 1, 2018, 3:07 p.m.
    ...acid sequence from within Snapgene, we can confirm the identity of the gene and check for any mutations...we identify the gene, we can easily annotate it by making a custom feature in Snapgene. This saves us ...frames, which is often our first clue about where the gene of interest might be located. By easily using BLAST...plasmids. We’ve also talked about how our new Snapgene generated maps provide improved feature detection with...’ll walk you through how a scientist at Addgene uses Snapgene to confirm the sequence of a plasmid and...through our Snapgene powered maps and sequence analysis tools.     The main ways we use SnapGene for plasmid...version of Snapgene to visualize the sequence and look for features. The first thing that the Snapgene software...

  8. Let There Be LITE Plasmids

    Type
    Blog Post
    Published
    Nov. 21, 2013, 3:56 p.m.
    ... after light stimulation. Under those optogenetic controls, gene expression levels reliably increased ...could be by applying it to altering gene expression via epigenetics with the addition of histone modifiers...conventional optogenetic tools." In total, the researchers applied the LITE system to 28 different genes to establish...transcriptional effectors (LITEs) designed to bind specific genes and turn them on or off, literally at the flip of...are able to react and learn. All of this requires genes to be regulated dynamically.” To understand how ... tools to manipulate the expression of specific genes in the brain in reversible fashion. And that’s exactly...demonstrated use of the LITEs to control gene expression in mouse neurons and in the brains of living mice ...

  9. An Interview with Connie Cepko - Gene Therapy, Plasmid Tools and Insights to Success

    Type
    Blog Post
    Published
    Dec. 8, 2015, 3:30 p.m.
    ...Strategies to Make Gene Therapies More General (more than correcting a defect in a single gene) 26:00-28:39 ...39 : Methods for Delivering Multiple Genes to Cells for Gene Therapy 28:40-35:10 : GFP Activatable Tools...Targeting in Retinal Gene Therapies 10:17-14:34 : Progress in Treating Macular Degeneration and Other Retinopathies...fluorescent proteins as scaffolds for cell-specific gene manipulation.” Cell 154(4) (2013): 928-939. PubMed... gene therapy...Production Specialist at Addgene. He is the audio engineer and producer for the Addgene Podcast.     References...plasmids from this publication at Addgene. Resources on the Addgene Blog Read our Recent Blog Post on...

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

    Type
    Blog Post
    Published
    June 28, 2018, 3:01 p.m.
    ... 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...

  11. 27 Hot Plasmids from 2016

    Type
    Blog Post
    Published
    Dec. 22, 2016, 3:03 p.m.
    ...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...

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

    Type
    Blog Post
    Published
    Dec. 10, 2020, 1 p.m.
    ...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...

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

    Type
    Blog Post
    Published
    Sept. 28, 2020, noon
    ...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...

  14. Fluorescent Proteins 101: Luciferase

    Type
    Blog Post
    Published
    June 24, 2014, 3:59 p.m.
    ...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 ...

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

    Type
    Blog Post
    Published
    June 17, 2021, 1:17 p.m.
    ...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...

  16. Viral Vectors 101: AAV Variables That Matter

    Type
    Blog Post
    Published
    June 1, 2023, 1:15 p.m.
    ...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) ...

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

    Type
    Blog Post
    Published
    July 23, 2019, 12:46 p.m.
    ...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...

  18. Viral Vectors 101: Producing Your rAAV

    Type
    Blog Post
    Published
    July 16, 2024, 1:15 p.m.
    ...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...

  19. Plasmids 101: Shuttle Vectors

    Type
    Blog Post
    Published
    May 20, 2025, 1:15 p.m.
    ...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...

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

    Type
    Blog Post
    Published
    May 9, 2019, 1:15 p.m.
    ...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...

  21. Meet the New Addgene Board Members!

    Type
    Blog Post
    Published
    April 28, 2020, 1:14 p.m.
    ...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...

  22. Hot Plasmids and Viral Preps - July 2021

    Type
    Blog Post
    Published
    July 27, 2021, 1:15 p.m.
    ...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...

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

    Type
    Blog Post
    Published
    April 13, 2021, 1:15 p.m.
    ...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 ...

  24. Viral Vectors 101: Integrase-Deficient Lentiviral Vectors

    Type
    Blog Post
    Published
    May 13, 2025, 1:15 p.m.
    ...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...

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

    Type
    Blog Post
    Published
    June 23, 2022, 1:15 p.m.
    ...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...

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

    Type
    Blog Post
    Published
    Oct. 17, 2019, 12:28 p.m.
    ...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...

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

    Type
    Blog Post
    Published
    Aug. 17, 2021, 1:15 p.m.
    ... 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...

  28. What's New in CRISPR - September 2019

    Type
    Blog Post
    Published
    Sept. 10, 2019, 12:56 p.m.
    ...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...

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

    Type
    Blog Post
    Published
    Aug. 6, 2019, 12:55 p.m.
    ...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...

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

    Type
    Blog Post
    Published
    Dec. 15, 2016, 3:57 p.m.
    ...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...

  31. PCR Applications for Copy Number Variation Assays

    Type
    Blog Post
    Published
    April 26, 2022, 1:15 p.m.
    ...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...

  32. Supporting Rare Disease Research with BeHeard 2017

    Type
    Blog Post
    Published
    July 20, 2017, 2:30 p.m.
    ...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 ...

  33. CASTing Off for New Shores in Human Genome Editing

    Type
    Blog Post
    Published
    June 10, 2025, 1:15 p.m.
    ...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...

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

    Type
    Blog Post
    Published
    Nov. 25, 2013, 10:12 p.m.
    ...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...

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

    Type
    Blog Post
    Published
    May 11, 2017, 2:30 p.m.
    ...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. ...

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

    Type
    Blog Post
    Published
    March 17, 2020, 1:32 p.m.
    ...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...

  37. Viral Vectors 101: Viral Vector Elements

    Type
    Blog Post
    Published
    July 25, 2023, 1 p.m.
    ...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!   ...

  38. Bacterial Toxin-antitoxin Systems as Molecular Biology Tools

    Type
    Blog Post
    Published
    Nov. 1, 2018, 12:35 p.m.
    ...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....

  39. Tips for Using FRET in Your Experiments

    Type
    Blog Post
    Published
    Nov. 5, 2014, 3:41 p.m.
    ...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...

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

    Type
    Blog Post
    Published
    Oct. 13, 2022, 1:45 p.m.
    ...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...
Showing: 81 - 120 of 920 results