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We narrowed to 251 results for: tre promoter

Showing: 61 - 80 of 251 results

  1. Targeting HIV-1 with CRISPR: Shock and Kill or Cut it Out?

    Type
    Blog Post
    ...repeat (LTR) of the HIV-1 genome, which acts as a promoter, and found that SAM targeting of the enhancer ...NF-KB binding sites increased activation of HIV promoter-luciferase constructs. In multiple HIV-1 latent...cells with activated HIV-1, measured via HIV-1 promoter-driven GFP expression. In cell lines that can ...indels may not ablate viral function, but rather promote resistance to CRISPR/Cas9. While these results ... acquired immunodeficiency syndrome (AIDS) in untreated individuals. Symptoms of AIDS include rapid weight... viral levels will soon skyrocket back to pre-treatment levels due to the viral reservoirs located in ...in latently infected cells. Although ART-treated patients lack plasma HIV-1, they are at increased risk...

  2. Harnessing Bacterial Toxins for Allelic Exchange

    Type
    Blog Post
    ...-inducible promoter, though Khetrapal et al. also demonstrated that the arabinose promoter worked too.... under both rhamnose- and arabinose-inducible promoters. Tight control of toxin expression is absolutely...a rhamnose-based tightly regulated and tunable promoter system." Biotechniques 40.3 (2006): 355-364. PubMed...routinely clone the purified amplicons upstream and downstream of the desired deletion directly into cut...transcriptional terminator BBa_B1002 (from IGEM) upstream of the MCS to reduce transcriptional readthrough...accessibility, selective RNA unfolding and sliding at upstream standby sites." Nucleic acids research42.4 (2013...

  3. Scientific Reproducibility - Focusing on Solutions at the Minisymposium on Reproducibility

    Type
    Blog Post
    ...resources and information about organizations promoting reproducibility in this booklet (which was also...line we may see further adoption of policies that promote reproducible publishing practices. One example ... also hopefully see a shift toward hiring and promotion practices that focus on reproducibility and ensuring... Including descriptions of many organizations promoting reproducibility List of Tools for Reproducibility...academia, industry, nonprofits, and publishing. The livestream video from the event can be found below along...interested in, but I’d recommend watching the whole livestream for a more holistic understand of reproducibility...reasons we alter these designs if we do so. The most extreme version of this (and possibly the most effective...

  4. Fluorescent Tagging of Endogenous Genes with SapTrap

    Type
    Blog Post
    ... site accepts the sgRNA target sequence for U6 promoter expression and the second site accepts the homology... a direct fusion of the desired gRNA to the U6 promoter, and a second plasmid containing the homology ...regions of homology to the gene of interest to promote homology directed repair after a CRISPR/Cas9 induced...T2A-TurboGFP-PEST), and small epitope tags (HA, Myc, Strep tag II, AviTag, HaloTag, SpyTag). The Foerstemann... N-terminal tagging vectors (eg GFP, Flag, YFP, Strep, TEV-V5) with either blasticidin or puromycin selection...

  5. Colony Formation Titering Assay

    Type
    Protocol
    .... Pro-Tip Different brands and lots of FBS can promote or inhibit transfection. Test a variety of brands...control the multiplicity of infection (MOI) in downstream studies. This protocol was developed for A549...Microcentrifuge tubes, Neptune 3745.X 6-well tissue culture treated dish, Corning 3516 15 mL conical tubes, VWR 21008... after 20–30 passages. Do not add penicillin/streptomycin to the media, as it can affect viability and...target cell line needs to be empirically determined. Treat the target cells with a range of doses of antibiotic...vector pRosetta . The following day, cells were treated with puromycin. Colonies were allowed to grow in...EF1α dCas9-KRAB . The following day, cells were treated with puromycin. Colonies were allowed to grow in...

  6. Hot Plasmids: Spring 2025

    Type
    Blog Post
    ...-shock proteins at the FOS gene promoter using TurboCas, streptavidin immunoprecipitation (IP), and tandem...new backbones special: Strong expression: A CMV promoter drives robust gene expression in mammalian cells...creation a breeze. Modular by design: Key elements (promoter, tag, polyA site) are flanked by unique restriction... both new and known regulators at the FOS gene promoter and the MYC locus (Figure 5). Figure 5:...

  7. Viral Vectors 101: Systemic Capsids

    Type
    Blog Post
    ...experiment is determined by the capsid and promoter/enhancer strengths in the cell type of interest as well ...transgene expression. Gene regulatory elements [promoters and enhancers, miRNA target sites, and recombination...over systemic delivery. Otherwise, cell-specific promoters and enhancers (Mich et al., 2021) can help restrict...optogenetic and chemogenetic tools. Dual expression Promoters, miRNAs, enhancers, and switches all take up precious...candidate regulatory elements such as enhancers and promoters for cell type-specific targeting (Chan et al.,...when paired with appropriate cell type-specific promoters or Cre lines. Spatial transcriptomics suggests...AAV-PHP.eB, when paired with the ubiquitous CAG promoter, is relatively biased toward L5 and inhibitory...

  8. Plasmid Cloning by Restriction Enzyme Digest (with Protocols)

    Type
    Protocol
    ...However, the same technique can be used to move promoters, selectable markers, or any other DNA element ...single enzyme, but this will require phosphatase treatment of your recipient plasmid as well as a specifically...re-circularization of the recipient plasmid. You should treat your digested recipient plasmid with a phosphatase...

  9. Isolating Genomic Regions of Interest with the CRISPR System

    Type
    Blog Post
    ... repression of transcription through binding a promoter region to inhibit association of transcriptional...targeting their genomic locus of interest, the promoter region of the transcription factor IRF-1. Immunoprecipitation...lab found a microRNA (miR483) that binds to the promoter of the growth factor IGF2 open reading frame to...dCas9, with bound DNA, can then be purified by streptavidin high-affinity purification. This system is both...

  10. Plasmids 101: Shuttle Vectors

    Type
    Blog Post
    ... actually express your gene of interest? Many promoters are species-specific, so you’ll need to design...However, you do still need a species-compatible promoter for expressing your selectable marker. Otherwise...Plasmids 101: Mammalian Vectors Plasmids 101: The Promoter Region Resources on Addgene.org Collection of...cells “speak” the same molecular language as the tree outside my window! Whatever you use them for, shuttle...

  11. Save Time with Transient Plant Leaf Transformations

    Type
    Blog Post
    ...are therefore good temporal reporters to measure promoter activity, for example. Useful positive controls...dark after the transformation. Use introns Plant promoters can be recognized or leaky in Agrobacterium. This...of the GUS gene with the commonly used CaMV 35S promoter in Agrobacterium. This can be easily avoided by...more easily quantify the effect of the different treatments on transformation success. Some reporter gene...

  12. CASTing Off for New Shores in Human Genome Editing

    Type
    Blog Post
    ...to a plasmid that lacked a promoter — only CAST integration of the promoter could turn the gene on. Once...integrate a payload sequence into genomic DNA downstream from the Cas target. Conveniently, the process...to integrate large sequences into genomic DNA downstream of gRNA-specified sites. Created with BioRender.com...pressures: high efficiency helps them spread, but also stresses the host. The researchers suspected natural evolution...emerging world of gene therapy, highly tailored treatments to specific mutations require individualized ... of loss-of-function mutants could provide a streamlined, one-size-fits-most approach. evoCAST is an important... important step toward that kind of gene treatment. The researchers also compared evoCAST to the Liu lab...

  13. Virus Protocol - Generating Stable Cell Lines

    Type
    Protocol
    ... °C. Pro-Tip Different brands and FBS lots can promote or inhibit transfection. Test a variety of brands...Microcentrifuge tubes, Neptune 3745.X 6-well tissue culture treated dish, Corning 3516 15 mL conical tubes, VWR 21008... after 20-30 passages. Do not add penicillin/streptomycin to the media, as it can affect viability and... reagent for your target cell line. To do this, treat target cells with a range of doses of antibiotic... try higher/lower dilutions depending on your downstream applications. If you’ve titered your virus beforehand...

  14. What Do I Do Now? Academic v. Non-Academic Career Decisions

    Type
    Blog Post
    ... researchers must be extreme self-starters and especially good at self-promoting—No one is going to tell...diversity and acceptance can be an issue in getting promoted (especially race and gender) Interpersonal issues...especially with peer mentors, to discuss your strengths, weaknesses, interests and passions is another...

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

    Type
    Blog Post
    ...limited by the lack of well characterized inducible promoters. Far removed from the bio-industry’s vision of... the same dCas9 protein. By using an inducible promoter to modulate dCas9 expression, multiple scRNA transcriptional...genes or to introduce heterologous genes, but the trend of research has shifted in recent years toward tinkering...regulators (Zalatan et al. 2015).  These scaffolds are stretches of non-coding RNA interspaced with linkers that...

  16. Addgene-Seeding Labs plasmid grant round 3: microbial drug resistance and biofilms, antidiabetic plant components, visualizing brain injury - and a glowing robot fish

    Type
    Blog Post
    ... sensor, he will construct plasmids containing promoter regions that activate fluorescent protein transcription...and to Seeding Labs who helped us coordinate and promote the awards. Additional Resources on the Addgene... has studied various aspects extreme life on earth including extremophile microorganisms, biomarkers, ... to pyrazinamide (PZA) (a drug commonly used to treat tuberculosis) and in identifying new drug targets...biofilm formation. The quest for plant compounds to treat complications of diabetes Gloria Otito Izu, PhD...infusions of L. taraxacifolia leaves are widely used to treat several diseases. Her goal is to identify and isolate...blocked, resulting in reduced blood flow. Novel treatment options, as well as non-pharmacological solutions...

  17. New and Upcoming Viral Vectors - May 2020

    Type
    Blog Post
    ...activated, this DREADD acts through the Gq pathway and promotes neuronal firing. Plasmid Serotype Name Depositor...more accurate cell and tissue targeting. While promoters and AAV serotypes can sometimes be used to target...2-N-2xStrep-IRES-Puro: Expression of the SARS-CoV-2 N protein  pLVX-EF1alpha-SARS-CoV-2-M-2xStrep-IRES-Puro...pLVX-EF1alpha-SARS-CoV-2-E-2xStrep-IRES-Puro) and NSP2 (pLVX-EF1alpha-SARS-CoV-2-nsp2-2xStrep-IRES-Puro) are also...SARS-CoV-2 M protein pLVX-EF1alpha-SARS-CoV-2-orf3a-2xStrep-IRES-Puro: Expression of the SARS-CoV-2 ORF3a ...

  18. Synthesized by Ginkgo Bioworks, Shared by Addgene: SARS-CoV-2 Plasmids for Many Expression Systems

    Type
    Blog Post
    ...expression plasmids in a high copy backbone with a T7 promoter driving multiple combinations of WT, codon optimized...which drive expression using either TEF1 or GAL1 promoters. These vectors are intended for the high level...race to find vaccines, therapeutics, and other treatments to stem the spread of COVID-19. That’s why we...sites (thrombin and TEV), and C-terminal tags (StrepII, 3xFLAG, 6xHIS, MBP, Myc). These constructs include...expression and purification of SARS-CoV-2 proteins for downstream experiments and offer flexibility in tag, cleavage...requested the plasmid. By leveraging our core strengths of intensive screening and the use of high throughput...

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

    Type
    Blog Post
    ...dCas9 along with transcriptional repressors to the promoter region of the gene of interest. dCas9-based repressors... and a dCas9-KRAB fusion, both targeted to the promoter region. Cas9 activators include Cas9 fused to ...the Suntag array of factors, all targeted to the promoter region.   Ok, I have a Cas9! What do i do ... genome.  2) The target is present immediately upstream of a Protospacer Adjacent Motif (PAM), the sequence...

  20. Multiple Plasmids at a Low Price: Kits from Addgene

    Type
    Blog Post
    ...different promoters followed by leader sequences (encoding small leader peptides) downstream of which ... vectors within this kit contain a variety of promoters to facilitate gene expression in bacteria, yeast...different genes was predictable based solely on the promoters and Shine-Delgarno sequences used in this so-called...transcription from resulting in the production of a downstream peptide, the transcription terminators in this...BIOFAB Plasmid Set contain terminators of varying strengths centered within GFP/RFP bicistronic reporters ...
Showing: 61 - 80 of 251 results