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We narrowed to 254 results for: tre promoter

Showing: 101 - 120 of 254 results

  1. PITChing MMEJ as an Alternative Route for Gene Editing

    Type
    Blog Post
    ...Addgene are perfect for expressing GFP from a given promoter, and the technique can be adapted to other transgenes...observed will be dependent on both the endogenous promoter and the 3’ UTR of the locus of interest, since...of a GFP-Puro cassette into a given locus, just upstream of a stop codon. Briefly, the PITCh vector should...since the GFP-Puro will be inserted just upstream of a stop codon. One potential concern is if the GFP-Puro...

  2. INTEGRATE: Bacterial Genome Engineering Using CRISPR-Transposons

    Type
    Blog Post
    ... VchINT, three separate plasmids and multiple promoters were used to express the necessary components ...required genes and the CRISPR RNA under the same promoter, and a donor plasmid (pDonor) encodes the mini-transposon...labs (Peters et al., 2017; Klompe et al., 2019; Strecker et al., 2019), which direct RNA-guided transposition...Halpin-Healy et al., 2020). Genomic targets require an upstream 2-bp PAM sequence for recognition by QCascade;...from the donor molecule, and inserts it ~50 bp downstream of the genomic target site bound by QCascade ...structures known as cointegrates (Vo et al., 2020; Strecker et al., 2020).  CRISPR-transposons for bacterial...et al., 2019). Recently, the lab introduced a streamlined expression approach (Fig. 2A), involving two-...

  3. Deciphering the Mysteries of Behavior with Viral Vectors

    Type
    Blog Post
    ... and triggering release of the stress hormone norepinephrine to promote vigilance. A recent study investigated... "Dorsal raphe neuroinflammation promotes dramatic behavioral stress dysregulation." Journal of Neuroscience...readout for associated emotional phenomena like stress levels, but researchers have to be mindful of motivation...coeruleus (LC) is known to play a major role in the stress response circuit by integrating neuronal signalling...freezing response, a form of immobility caused by stress and fear. Such a behavior can be monitored in an... 1970s to test the efficacy of antidepressant treatment in rodents. In this test, an animal is placed ... cortical noradrenergic excitability upon acute stress." The EMBO journal 37.21 (2018). PubMed PMID: 30209240...

  4. Addgene’s Top 10 Blog Posts from 2014

    Type
    Blog Post
    ...Competent Cells By Melina Fan Plasmids 101: The Promoter Region – Let's Go! By Kendall Morgan Plasmids...the experts in the CRISPR/Cas field, have been extremely helpful for scientists that are looking for more...on “Making Your Own Competent Cells” has been extremely popular, generating many additional questions....

  5. A New Generation of Adenine Base Editors Improves Editing in Primary Human Cells

    Type
    Blog Post
    ...studies have shown that one base change in the promoter region of gamma hemoglobin (HGB1/2) leads to continuous.... ABE8 efficiently edited the fetal hemoglobin promoter and caused persistent expression of fetal hemoglobin... domain can edit the short single-stranded DNA stretch that is accessible. Image from Gaudelli et al.,...

  7. Chromoproteins: Colorful Proteins For Molecular Biology Experiments

    Type
    Blog Post
    ... toxin gene (regulated by a rhamnose-inducible promoter) and a chromoprotein gene. Allelic exchange occurs...it is under the control of a rhamnose-inducible promoter. The resolution step leads to the desired mutation... the loss of their symbiotic algae, during heat stress. For other organisms, like the sea anemone, scientists...

  8. Lentiviral Vector Uses and Overview

    Type
    Blog Post
    ...a chimeric 5’ LTR containing a Tat-independent promoter. Although safer, this system may be more cumbersome... vectors integrate into the genome, they could promote oncogenesis by altering local gene expression. ...HIV-1 envelope. As of 2013, 65 patients had been treated with this therapy, which lowered viral load and...15(2):78-82. PubMed PMID: 23322669. 6. Genetic treatment of a molecular disorder: gene therapy approaches...

  9. Tips for Screening with Yeast Two Hybrid Systems

    Type
    Blog Post
    ...that also contains a reporter gene cloned downstream of a promoter containing the target sequence of the ...as the “prey”) in close enough proximity to the promoter of the reporter gene to activate expression (Figure...fusion protein shown is still able to bind to the upstream activation sequence (UAS) of its target gene, ...

  10. Bacterial Expression Systems

    Type
    Collection
    ...coli strain. Addgene Blog Inducible Promoters Repressible Promoters Synthetic Photobiology: Optogenetics...pRU1701 Promoter activity Fluorescence (GFP+) Gram-negative bacteria Philip Poole 14460 pOT2 Promoter activity...pCHERRY3 Promoter activity Fluorescence (mCherry) Mycobacterium sp. Tanya Parish 24657 pASTA3 Promoter activity...#1000000037) contains specific bacterial promoters that vary in strength and that can be used to reliably drive...periplasmic localization: PelB, MalE, OmpA ID Plasmid Promoter Tags PI Additional Addgene Protein Purification...Toolkit (Addgene #1000000080) features a library of promoters and purification tags compatible with Golden Gate...temperature, in different bacterial systems. ID Plasmid Promoter Inducer Expression Species PI 44249 pdCas9-bacteria...

  11. Bacterial Toxin-antitoxin Systems as Molecular Biology Tools

    Type
    Blog Post
    .... MazF expression is under control of the AOX1 promoter whose expression is induced by methanol. Once ...luciferase-expressing plasmids to examine in vivo-induced promoters in the Vibrio cholerae vaccine strain CVD 103-...However, the sok antisense-RNA binds a sequence downstream of the mok SD resulting in a double stranded ...

  12. Synthetic Biology, Artificial Chromosomes, iGEM & More - Interview with Tom Ellis

    Type
    Blog Post
    ...screening direct interactions between drugs and promoters of oncogenes. He returned to academic research...Podcast features guest host and Addgene European Outreach Scientist, Benoit Giquel. Traveling from our offices...in Academia. Will it all pay off? Chapter 2: The Centre for Synthetic Biology and Innovation - A shared...Tom Ellis! Dr. Ellis is a lead researcher at the Centre for Synthetic Biology and Innovation, Imperial ...project. His team also investigates bacteria and extremophiles for biotechnology and has been directly involved...

  13. The Fluorescent Vegetables in Aptamer Soup

    Type
    Blog Post
    ...transcription by placing the aptamer downstream of a known promoter. Addgene’s Plasmids 101: Aptamer Fluorophores... RNA Polymerase III. By expressing Corn under promoters for transcripts uniquely transcribed by RNA Polymerase...for imaging in live cells. By placing Spinach downstream of a riboswitch, metabolite binding to the riboswitch...ability to track RNA dynamics in a model of Fragile-X tremor/ataxia syndrome (Strack et al., 2013). Since then...

  14. CRISPR Activation: A Practical Guide

    Type
    Blog Post
    ...Cas9 would have an off-target that lands in the promoter of another gene, thereby driving aberrant transcription... said, we generally pick guides by putting the promoter of the gene into a gRNA finder such as WU-CRISPR...targeting the guides to a region less than 200 bp upstream of TSS for best results but up to 400 bp works...10.1016/j.cell.2014.09.029 Konermann S, Brigham MD, Trevino AE, Joung J, Abudayyeh OO, Barcena C, Hsu PD, Habib...

  15. Adeno-associated Viruses (AAVs) for Genome Editing

    Type
    Blog Post
    ...the targeting vector is inserted downstream of a functional promoter (gene traps). The resultant gene ...pAAV-TK-Acceptor has a conventional heterologous promoter-driven TK-neoR gene. To modify these tagging vectors...pAAV-SEPT-Acceptor, the preferred vector, contains a promoterless splice acceptor-IRES-neoR gene, which enhances...AAV preps generally used for gene editing are low titre, and are therefore able to infect only a small subset...

  16. CRISPR/Cas9 FAQs Answered!

    Type
    Blog Post
    ...sequence into a backbone that uses the human U6 promoter to drive expression, is it necessary to add a ... start of my target sequence? A3: The human U6 promoter prefers a 'G' at the transcription start site ...of the crRNA) is also expressed from a separate promoter. In the pre-processing form, the crRNA contains...they also be targeted? A4: The Cas9 cuts 3–4 bp upstream of the PAM sequence. There can be some off-target...suggestions? A14: The Zhang lab recommends using the Epicentre QuickExtract solution to perform this step to ...D., Lin, C., Gootenberg, J. S., Konermann, S., Trevino, A. E., Scott, D. A., Inoue, A., Matoba, S., Zhang...

  17. Plasmids 101: Common Lab E. coli Strains

    Type
    Blog Post
    ...your plasmid – you carefully selected the optimal promoter for your gene of interest, painstakingly cloned...Plasmids 101 Posts: Origin of Replication The Promoter Region – Let's Go! Mammalian Vectors Looking ..., and may have even put a fluorescent protein downstream, separated by an IRES element. You did a lot ...subunit S12 of 30S ribosome Confers resistance to streptomycin Tn10   Confers resistance to tetracycline ...galK rpsL (StrR) endA1 nupG fhuA::IS2 DB3.1 Streptomycin  HB101 derivative. For propagating plasmids ...lacY1 proA2 rpsL20(Smr) xyl5 Δleu mtl1 DH10B Streptomycin MC1061 derivative. General cloning and storage...lacZYA-argF)U169, hsdR17(rK- mK+), λ– HB101 Streptomycin Hybrid of E. coli K12 and E. coli B (mostly ...

  18. Twenty Years of Sharing: Expanding Accessibility

    Type
    Blog Post
    ... pages, we added curation fields for purpose, promoters, expression types, and more, all of which help...How to Design Your gRNA for CRISPR Editing The Promoter Region - Let's Go! How to Verify Your Plasmid ... easier access to export-controlled items by streamlining permissions and paperwork. Through updating ...

  19. Plasmids 101: Broad Host Range Plasmids

    Type
    Blog Post
    ...resistance cassettes (Prior et al. 2010), inducible promoters with different degrees of stringency and inducibility...Development of constitutive and IPTG-inducible integron promoter-based expression systems for Escherichia coli ...spheroides, Salmonella spp., Serratia marcescens, Streptomyces lividans, Synechococcus spp., Thiobacillus ferrooxidans...

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

    Type
    Blog Post
    ...following changes. In pCas9, we 1) replaced the promoter of gRNA-pMB1 with PrhaB, 2) changed the replicon...pCutamp cures the plasmids by targeting the ampR promoter in all three plasmids. pCutamp is then removed... B strains, 1 E. coli W strain, and 1 Tatumella citrea, with a total of 70 sites edited. We optimized ...
Showing: 101 - 120 of 254 results