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We narrowed to 130 results for: tac promoter

Showing: 1 - 20 of 130 results

  2. Plasmids 101: Repressible Promoters

    Type
    Blog Post
    ... are repressible promoters regulated? Like inducible promoters, repressible promoters can be regulated...Mark Vasser. “The tac promoter: a functional hybrid derived from the trp and lac promoters.” Proc Natl Acad...Promoters control the binding of RNA polymerase and transcription factors. Since the promoter region ...natural and engineered repressible promoters. Chemically repressible promoters Tet-Off In its native context... pADH1 displays low promoter activity (~20% activity of the strong yeast promoter pTEF). As ethanol accumulates...that promoter is active. In this way, you can interrogate the activity of uncharacterized promoters. Similarly... QUAS promoter, and the QF transcription factor cannot drive transcription from the UAS promoter. This...

  3. Plasmids 101: The Promoter Region – Let's Go!

    Type
    Blog Post
    ... trying to pick your perfect promoter! Eukaryotic Promoters   Promoter   Primarily used for RNA     .... In practice, the term "promoter" describes the combination of the promoter (RNA polymerase binding site...recognize and bind to specific promoter elements. This means that the promoter present in your plasmid backbone...RNAP II promoter, whereas small RNAs (such as shRNA) are transcribed from the RNAP III promoters. This ...post on viral vector parts. Promoter specificity Aside from choosing a promoter based on type of RNA transcript...cell types or organisms, promoters must be similarly variable. Bacterial promoters only work in prokaryotic...require unique promoters and there is very little crossover. Generally speaking, promoters in bacteria are...

  4. Neurodegeneration Plasmid Collection

    Type
    Collection
    ...Yeast Other Promoter CMV T7 polH GAL Other Clear Filters ID Plasmid Name Gene Tags Promoter Disease PI ...PINK1 WT PINK1 GST tac Parkinson's Mark Cookson 13322 pGEX5X.1 PINK1 KD PINK1 GST tac Parkinson's Mark ...DJ1-WT PARK7 GST tac Parkinson's Mark Cookson 29351 pGEX-5X-1-DJ1-D149A PARK7 GST tac Parkinson's Mark...DJ1-E64D PARK7 GST tac Parkinson's Mark Cookson 29353 pGEX-5X-1-DJ1-M26I PARK7 GST tac Parkinson's Mark ...44978 GST-FUS-WT FUS GST tac ALS Robin Reed 44981 GST-FUS 266-526 FUS GST tac ALS Robin Reed 44982 GST-FUS...GST-FUS 356-526 FUS GST tac ALS Robin Reed 44983 GST-FUS 465-526 FUS GST tac ALS Robin Reed 44984 GST-FUS...pGEX-parkin WT PRKN GST tac Parkinson's Kalle Gehring 45970 pGEX-parkin 77-465 PRKN GST tac Parkinson's Kalle...

  5. Plasmids 101: Modular Cloning

    Type
    Blog Post
    ... individual part, this category also includes promoters, UTRs, terminators, etc. Once selected, individual...one or more gene(s), and typically begin with a promoter and end with a terminator. A simple TU could be... be comprised of just three parts: promoter, gene, and terminator.     Fig 1. MoClo enables individual...three steps: Level 0: Cloning of basic parts (promoters, UTRs, genes, tags, etc.) into MoClo-compatible...maximize production potential. For example, the best promoter for a given peptide can easily be tested by generating...generating Level 1 TUs with unique promoters, and the ideal phosphatase (or any modifying enzyme) to pair...additional components such as selectable markers, promoters, enhancers, etc. MoClo is especially useful for...

  6. CRISPR 101: Multiplex Expression of gRNAs

    Type
    Blog Post
    ...from its own promoter. But that doesn’t mean you have to clone and transfect multiple promoter-gRNA constructs...can I express more than one gRNA from a single promoter using a plasmid like pX330? Unfortunately, the...of pX330, contains humanized wtCas9 and two U6 promoters. To use this plasmid, you simply order oligonucleotides...contain Type IIS restriction sites flanking the promoter-gRNA construct, but with different sequences adjacent...contain Type IIS restriction sites that flank the promoter-gRNA constructs. When these plasmids are digested...downstream of the 7SK, human U6, mouse U6, or human H1 promoters. If you express fewer than four gRNAs, you’ll ...in a polyT-termination sequence for each unused promoter. This step is necessary to generate all of the...

  7. Recombinase-based State Machines Enable Order-dependent Logic in vivo

    Type
    Blog Post
    ...inducible promoters: the BxbI gene downstream of the anhydrotetracycline (ATc)-inducible promoter (PLtetO...and promoters with buffer sequences on their 5’ and 3’ ends (for insulation). A list of promoters and ...gene downstream of the arabinose (Ara)-inducible promoter (PBAD), and the A118 gene downstream of the diacetylphloroglucinol...diacetylphloroglucinol (DAPG)-inducible promoter (PPhlF).  Output Plasmid: The output plasmid (single...Depending on the desired application, the inducible promoters we used to drive recombinase expression may be...demonstrates that genetic parts (genes, terminators, and promoters) may be interweaved onto a GRSM register to execute...in this paper (recognition sites, terminators, promoters, and genes) were built with flanking primer binding...

  8. A Practical Guide to Optimizing AAV DIO and FLEx Vector Expression

    Type
    Blog Post
    ...the injection site. Test the promoter It is also essential that the promoter be thoroughly tested in your...cell-type specific” promoter, such as the truncated tyrosine hydroxylase (TH) promoter for catecholamine...open reading frame is inverted in relation to the promoter, hence the name, double-floxed inverted open-reading...what can we do about it?  Test the serotype and promoter The first step is to know how the AAV vector you...includes a thorough testing of both the serotype and promoter with in vivo experiments to determine the optimal...your cells.  Some promoters, such as CAG are very strong and may require less viral particles or relatively...expressing TH protein indicating that there is promoter activity. Additionally, some neurons express low...

  9. The Breast Cancer Microenvironment: A Tumor’s Backstage Team

    Type
    Blog Post
    .... Unfortunately, as a whole, microenvironments promote tumor durability instead of repressing them, making...a look at cancer cell metabolism. Cancer cells promote oxidative stress leading to the degradation of ...caveolin 1, also known as CAV 1. Usually, caveolin 1 promotes the cell life cycle progression and enhances tumor...signaling, possibly inducing the secretion of growth promoting proteins, such as human growth factor, vascular...Fibroblasts, or CAFs. CAFs are altered fibroblasts and promote the development and metastasis of tumors through...different organs. This means that exosomes not only promote microenvironment’s malignancy but also incite different...all, the system built to protect us now seems to attack us by releasing mitogenic growth mediators and ...

  10. Hot Plasmids - December 2019 - CRISPR pooled library, LipoGlo, CRISPRa, and Voltage Imaging Tools

    Type
    Blog Post
    ...σ54-based transcription. Transcription from σ54 promoters requires binding of bacterial enhancer binding...proteins (bEBPs) at a sequence upstream of the promoter called the upstream activating sequence (UAS)....gRNA complex which allows transcription from the promoter.  They found that the system is active in E. coli... and displays high dynamic range. However, the promoter in question must be σ54-dependent and the system...and further the study of natural σ54-dependent promoter systems. Find these CRISPR activation plasmids...information. Such voltage imaging can be performed in intact animals and offers many advantages over traditional...

  11. Viral Vectors 101: Parts of the AAV Packaging Plasmid

    Type
    Blog Post
    ...expression is often driven by the AAV p5 or p19 promoters to, respectively, produce the large (Rep78 and... the transcriptional regulation of the AAV p40 promoter which drives Cap expression. Often Rep proteins...genome. The cap gene is often driven by the AAV p40 promoter. Alternative splicing and differing translational...alternative open reading frame (ORF). While AAP promotes virion assembly, the role of MAAP has not been... domain. Following replication, Rep78/68 stays attached to the AAV genome and docks with the genome onto...

  12. Plasmids 101: The Wide World of Natural Plasmids

    Type
    Blog Post
    ...Electroporation is one popular method scientists use to promote transformation, but it isn’t just for lab specimens...specimens — there is evidence that lightning can also promote gene transfer in soil bacteria! (Demanèche et al...chromosome, but when encoded on plasmids, they can promote rapid spreading of genetic material through a population... features, such as origins of replication and promoters, check out our other Plasmids 101 posts, and keep...genes that help bacteria invade hosts, including attaching to host cells and evading the host immune system...resistance plasmids, which protect the host from attacks by other organisms. To conserve energy, these specialized...genetic material to a recipient through direct contact. That’s conjugation! F factors can also be integrated...

  13. 28 Hot Plasmid Technologies from 2015

    Type
    Blog Post
    ...fluorescent proteins, 8 constitutive promoters, 2 includible promoters, 3 polyA terminators and various pieces...sequences. Several pre-constructed promoter-selection marker or promoter-inducible expression related vectors...all chemically inducible promoters; 3) improving the strength of the promoters used; and 4) optimizing ...ideal promoter/location combination. However, other constructs which contain different promoters and/or...to study the human GM-CSF promoter and enhancer, a finely regulated promoter controlled by a mixture of...multiple copies of a transcription factor to a promoter and localization of a protein via the presence...are inserted between the truncated gene and the promoter to prevent leaky expression of this resistance...

  14. Experimenting with New Careers while in Grad School

    Type
    Blog Post
    ...career in science outreach: engaging the public to promote an awareness and understanding of science. Like...spray paint to make a trophy for the winner. We promoted the contest for about two months, and we got one...social media. I gained a concrete example to use to promote future attempts and learned a bit about recruiting...network and graduate school resources daily to promote CINSO events and recruit speakers and participants...educating the public and organizing events. I contacted local professional organizations and nonprofits...Science Indiana and sent emails to any group with a “Contact Us” button on their website. Then I had a lot of...Without the encouragement and pep talks from these contacts, I would have quit my “nontraditional” career ...

  15. Plasmids 101: Cre-lox

    Type
    Blog Post
    ...examined a number of important plasmid elements – promoters, origins of replication, protein tags, and antibiotic...Regulated Cre expression – placing Cre downstream of promoters that are active only in certain cell or tissue...Cre under a variety of ubiquitous and regulated promoters, and many loxP-containing alleles have also been...sequence. Among these are loxN (GtATACcT), lox2272 (GgATACtT), and lox511 (GtATACAT). These variant lox sites...canonical loxP sequence is ATAACTTCGTATA-GCATACAT-TATACGAAGTTAT. The loxP sequence does not occur naturally...

  16. AAV Production in HEK293 Cells

    Type
    Protocol
    ...) Pro-Tip Different brands and lots of FBS can promote or inhibit transfection. Test a variety of brands...viscosity of PEG. Stirring under medium heat will promote faster dissolution. Adjust the pH to ~7.4. Autoclave...flask, Corning 430825, 175 cm 2 Cellstack 5, Corning 3319, 3180 cm 2 Cellstack 2, Corning 3269, 1272 cm 2 ...-3 minutes for cells to detach. Gently tap the sides of the CS2 to help detach the cells, add 200 mL of...used to produce AAV from one Five Chambers Cell-Stack (CS5) (Link opens in a new window) (3,180 cm 2 -...the same surface area as 21 x T-175 flasks). Cell stacks provide an efficient means to scale-up without ...Pro-Tip While adherent, these cells are very loosely attached to the dish surface and should be handled carefully...

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

    Type
    Blog Post
    ...reducing off-target effects since promoter and enhancer sequences remain intact (Engreitz et al. 2019). The ...transcription start site, splice sites, exons, and/or promoters to knock out lncRNA function (Hazan and Bester...of introns or the removal of exons. Disrupting promoter or transcription start site function would also...Local regulation of gene expression by lncRNA promoters, transcription, and splicing. Nature. 2016;539...

  18. Introducing Addgene's New Search - Find Plasmids More Easily For Your Research!

    Type
    Blog Post
    ...scientists Viral prep filter categories Promoters: Select the best promoter for your target cell type Serotype...world, it can be hard to find your favorite gene, promoter, serotype, or a myriad of other things. That’s...plasmid to express ubiquitin under control of a CMV promoter. Starting with two search terms we see there are...coming up with no results at all, let us know! Contact our scientific technical support at [email protected]...

  19. 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 ...how CRISPR can take on HIV-1 in vitro, and what obstacles must be overcome for clinical success. ART and...ART-induced viral control found in most HIV-1 patients. Obstacles for CRISPR HIV-1 therapy Both of these methods...

  20. 27 Hot Plasmids from 2016

    Type
    Blog Post
    ... gRNA from a Drosophila U6:2 promoter and Cas9 from the actin 5C promoter. Addgene’s fly library web page... over 30 promoters and 140 genes, including constructs with unique tissue-specific promoters expressing...genomic libraries are cloned downstream of a minimal promoter sequence. Should one of these random stretches...activate its own transcription from the minimal promoter and its strength can be determined from its enrichment...variety of plasmids containing many different promoters, RBS’, coding sequences (CDS), and terminators...fluorophores, generate expression vectors with various promoters suitable for in vivo expression, and/or produce...continuous expansion of compatible backbones, promoters, and genes available to the community. If you ...
Showing: 1 - 20 of 130 results