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Showing: 81 - 100 of 244 results
  1. AAV Q&A with Tim Miles

    Type
    Blog Post
    ...efficient AAV prep for a 4.2 kB insert with the CMV promoter using the AAV-BR1 capsid? A: It is our understanding...4.7kB.  If your insert alone is 4.2 kB and CMV promoter+enhancer is 0.5 kB, then you would be oversized... reliable quantitation. Differences in sample treatment across methods can affect reported titer by either...
  2. Behind the scenes: Addgene’s new search engine and more

    Type
    Blog Post
    ...search scoped to viral preps shows filters for promoter and serotype, among others. We also created a ...example, there is only one viral prep filter for promoter “Syn”, that, when selected, will match variations...members from the Customer Service, Data Curation, Outreach, Product, Scientist, and Software teams. We knew...
  3. Evolution of Brainbow: Using Cre-lox for Multicolor Labeling of Neurons

    Type
    Blog Post
    ...protein whose coding sequence is closest to the promoter will be expressed. This system differs from Brainbow...translation-blocking STOP cassette immediately following the promoter. The STOP cassette includes a mutant YFP that .../FRT recombinase. When placed under different promoters, Brainbow and Flpbow can be used to label distinct...Drosophila. Why Brainbow? Brainbow highlights the trend in biology towards single-cell resolution imaging...
  4. Plasmid Tools for Microbiome Studies

    Type
    Blog Post
    ...suitability of various antibiotic resistance markers, promoters, and inducible gene expression systems in bee ...fusion is under control of the arabinose-inducible promoter. Cells are first induced with arabinose to begin...designed to bind and detect nearly any RNA sequence. Downstream of the hairpin is a ribosome binding site (RBS...responds to infection, dysbiosis, and antibiotic treatment. To facilitate these studies, Pamela Silver’s ...
  5. Working with Nuclear Receptors

    Type
    Blog Post
    ... Atlas (NURSA), one of our primary goals is to promote an understanding of how these processes are regulated...upon a context defined by ligand, cell type, and promoter, organizing the expression of functional groups...transported from cells, traverse the body through the bloodstream, and eventually elicit their actions on other...drugs currently available or being developed to treat a multitude of therapeutic indices, including hypertension...
  6. 22 Hot Plasmid Technologies from 2014

    Type
    Blog Post
    ...either the Thy1 or CAG promoter, while Brainbow AAV is under control of the EF1a promoter. Livet et al., ...known as SpyLigase and is a protein domain that promotes the formation of an isopeptide bond between 2 ...insert fragments of DNA containing basic parts (promoters, UTRs, coding sequences, terminators, etc) into... creating a single transcriptional unit (Ex: a promoter, 5’UTR, coding region, and terminator). Next, ...individual types of pre-cloned insert modules (plant promoter, N-terminal tag, coding sequence of the gene of...Biotech. 2013. Dec 23.   Highly bright and extremely dim GFPs The availability of the three dimensional...unprecedentedly-high brightness, bfloGFPa1, and the other with extremely-dim brightness, bfloGFPc1. The plasmids used for...
  7. 15 Years of Addgene: The Top 15 Plasmids

    Type
    Blog Post
    ...customizable sgRNA from the U6 promoter and puromycin resistance from the EF-1a promoter. It’s a 3rd generation...plasmid from the Zhang lab expresses Cas9 from Streptococcus pyogenes with an invariant sgRNA scaffold and...
  8. Hot Plasmids - March 2020 - base editors, GEVI, MoClo, and optogenetics

    Type
    Blog Post
    ...kit contains 96 parts including terminators and promoters, CRISPRi parts, selectable markers, transformation...determined protein expression levels from different promoters, and inhibited gene expression using CRISPRi. ... acceptor vectors from the MoClo Toolkit from Sylvestre Marillonnet’s lab to develop synthetic biology...
  9. Mouse Modeling, Part 1: Genetically Engineered Mice

    Type
    Blog Post
    ...vector containing homologous ends to the breakpoint promotes homologous recombination of the targeting vector... process. The addition of small molecules that promote homologous recombination or the use of long single-stranded...recessive traits and their impact on diseases or treatments, but are not useful for genetic engineering. ...initially inhibited by the presence of a stop cassette upstream of the target gene in the targeting vector. The...
  10. Delivery Methods for Generating iPSCs

    Type
    Blog Post
    ...expresses multiple reprogramming factors driven by one promoter. While lentivirus has similar rates of reprogramming...control. Additionally, Sendai virus doesn’t rely on promoters for transcriptional regulation, so it requires... mini-plasmids. They contain only a eukaryotic promoter and the cDNA(s) to be expressed and they don’t...phosphatase, expression of embryonic proteins, and promoter demethylation and expression of endogenous pluripotency... cellular defenses against ssRNA: phosphatase treatment, substituting cytidine with 5-methylcytidine, ...
  11. Teaching an Old DOG New Tricks: Controlling Protein Activity with GFP

    Type
    Blog Post
    ...very realistic. Envisioning GFP as a substrate promoting dimerization, Tang et al. tested pairs of their... they activate gene expression from different promoters. Moving to an in vivo system, Tang et al. electroporated...at LoxP sites. Placing a lox-STOP-lox cassette upstream of a gene of interest blocks transcription when...
  12. Viral Vectors 101: Calcium Sensors

    Type
    Blog Post
    ...neuron-specific expression (hSyn promoter) and pan-mammalian cell expression (CAG promoter). An improved version ...distinct cell subtypes for efficient expression and downstream monitoring. This targeted approach allows scientists...
  13. Mobile-CRISPRi: Bringing CRISPRi to Diverse Bacteria

    Type
    Blog Post
    ...allows for customization with strain-specific promoters, antibiotic resistance selection markers, gRNAs...nature allows for the customization of guides, promoters, selection markers, and different versions of ...trimethoprim when compared to control gRNA treated or untreated bacteria. Had the folA gene product not been..., Oren Rosenberg, and Carol Gross provides a streamlined suite of tools for using CRISPRi in many types...tri-parental mating scheme. Mobile-CRISPRi integrates downstream of the highly conserved glutamine-fructose-6-...and right Tn7 end sequences (green) integrates downstream of the glmS gene in the recipient's genome. Antibiotic...were grown in presence of trimethoprim, cultures treated with a folA gRNA were more sensitive to trimethoprim...
  14. Interview: Nicola Patron on Plant Synthetic Biology, MoClo, and More

    Type
    Blog Post
    ...plant lab, so it includes everyone’s favorite promoters, plus a few new ones that are not used so much...know how to start. They might want to use our promoters and terminators, but then they aren’t sure how...motivates her research, the MoClo Kit she and Sylvestre Marillonnet share with the scientific community...with the Athena SWAN Charter and it has really strengthened the network of female scientists. Twitter has..., Jonathan D. G. Jones, Nicola J. Patron, and Sylvestre Marillonnet. ACS Synth. Biol., Just Accepted Manuscript...
  15. Pathways Over Time Plasmids Engage Students in Functional Genomics Research

    Type
    Blog Post
    ... the pYES2.1 plasmid is controlled by the GAL1 promoter. The cloning procedure adds both V5 and His6 epitope...grow in the absence of methionine. Since the GAL1 promoter controls MET/Met gene expression, complementation...Associate Professor at Boston College. National reports stress the importance of providing authentic research ...
  16. 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...
  17. 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-...
  18. 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...
  19. Protocol - pLKO.1 – TRC Cloning Vector

    Type
    Protocol
    ...10878/ . Description Vector Element U6 Human U6 promoter drives RNA Polymerase III transcription for generation...transduced cells. hPGK Human phosphoglycerate kinase promoter drives expression of puromycin. Puro R Puromycin...with effective gene silencing: Starting at 25nt downstream of the start codon (ATG), search for 21nt sequences...position 15-19. Avoid targeting introns. Avoid stretches of 4 or more nucleotide repeats, especially repeated...Bovine Serum (FBS) Invitrogen: #16000 Penicillin/Streptomycin Invitrogen: #15140-122 Polypropylene tubes VWR...be passaged in DMEM + 10% FBS with penicillin/streptomycin, cells should be plated at this step in DMEM...10% FBS without antibiotics (no penicillin or streptomycin). Day 2: b. Perform the transfection in the ...
Showing: 81 - 100 of 244 results