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We narrowed to 356 results for: SAM activators

Showing: 81 - 100 of 356 results

  1. Using Addgene's New Viral Service for Your Research

    Type
    Blog Post
    ...delivery of Cas9 endonuclease, Cas9 nickase, Cas9 activators, Cas9 repressors, select gRNAs, and CRISPR pooled...variety of experiments, while rtTA can be used to activate expression of a gene under the control of the ...available allowing you to chemically control neuronal activity while studying your phenotype of interest. Optogenetic...Optogenetic tools for similarly controlling neuronal activity with light will be available soon. We’re also ...titering, and testing viral preps for them. At the same time, we’re developing ourselves as a knowledge ...

  2. Hot Plasmids April 2018 - Protein Degradation, Nanoscopy, FIRE-Cas9, and Yeast Expression Tools

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    Blog Post
    ...binding domains and the MSN2 transactivation domain. Transcription factor activation via the appropriate ligand...that TRIM21 overexpression does not aberrantly activate the proteasome. Clift et al. tested multiple methods...variety of protein modules to repress or activate gene expression, sometimes incorporating chemically inducible...binding to the appropriate promoter sequence and activation of gene expression. In addition to plasmids encoding...study epigenetic modulation. While catalytically inactive cas9 (e.g. dCas9) has been fused to a variety ...Factors Podcast Segment Researchers from the El-Samad lab recently deposited plasmids encoding synthetic...encoding these synthetic transcription factors, the El-Samad lab has deposited plasmids encoding fluorescent ...

  3. When Fidelity Matters: A frank discussion about ligase fidelity

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    Blog Post
    ...and is active over a range of elevated temperatures (37–75 °C). T4 DNA ligase is highly active in nick... applications, there cannot be any end-joining activity, and for others, there is a need for the exclusive...ligate mismatches to a significant degree, and very active ligases, such as T4 DNA Ligase, can ligate nicks...and A:C mismatches. Thermostable DNA ligases are active at elevated temperatures, allowing further discrimination...combination of the intrinsic selectivity of the ligase active site and careful balance of reaction conditions...Bruce Wallace. "Specificity of the nick-closing activity of bacteriophage T4 DNA ligase." Gene 76.2 (1989...Watson-Crick base pair, are joined with nearly the same efficiency as a correct base pair. DNA ligases have...

  4. Intro to Our Summer SciComm Series!

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    Blog Post
    ...practices in the field, and even getting into some active science communication research.   This summer,... really, the possibilities are endless.    Activity Example audience  Talking about your career...defining your audience for science communication activities  Many people are concerned that by defining a...good communication is making sure everyone’s on the same page. So for our first post, I thought it best to...communication by and to scientists in (roughly) the same field, often through manuscripts, seminars, or posters...PCR test” to mean “a test that uses a biological sample to look for presence of a virus.” A scientist would...consistent and clear.   And now that we’re all on the same page, we’re ready to get into the more exciting ...

  5. Hot Plasmids - June 2020 - Barcoded CRISPR Library, Sparse Cell Labeling, Calcineurin Reporter, and DNA Staining Dye Alternative

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    Blog Post
    ...contributed by Aliyah Weinstein Sparse Predictive Activity through Recombinase Competition (SPARC) is a fully...788679 A destabilized GFP reporter for calcineurin activity in yeast Article contributed by Leah Schwiesow...propidium iodine (PI), allowing for calcineurin activity measurements in stressed or aging cell populations...al., 2020. Find the GFP reporter calcineurin activity here Diessl et al., Microbial Cell, 2020. https...performance of each gRNA multiple times within the same experiment by counting both the gRNAs and their ...required for virulence in pathogenic fungi, it is an attractive drug target. Calcineurin reporters allow scientists...

  6. Immunology Research Plasmids and Resources

    Type
    Collection
    ...PLAU plasminogen activator, urokinase ATF, UPA, URK, u-PA PLAUR plasminogen activator, urokinase receptor...mitogen-activated protein kinase kinase 1 MAPKK1, MEK1, MKK1, PRKMK1 MAP2K2 mitogen-activated protein ...mitogen-activated protein kinase kinase kinase 14 FTDCR1B, HS, HSNIK, NIK MAP3K8 mitogen-activated protein...Cdc42/Rac)-activated kinase 1 MGC130000, MGC130001, PAKalpha PAK2 p21 protein (Cdc42/Rac)-activated kinase... protein (Cdc42/Rac)-activated kinase 4 - PAK6 p21 protein (Cdc42/Rac)-activated kinase 6 PAK5 PAK7 p21...the immune system triggers several processes to activate cells to recognize and get rid of infectious pathogens...

  7. Tagging Optogenetics and Chemogenetics Receptors: Fluorescent Proteins and Other Options

    Type
    Blog Post
    ...chemogenetics are powerful tools to modulate the activity of neurons and other brain cells. Since the opsins...receptor, one of the Designer Receptor Exclusively Activated by Designer Drugs (DREADDs) (Armbruster et al....need to be located on the plasma membrane to be activated by the DREADD actuator after it has been administered...family of G protein-coupled receptors potently activated by an inert ligand. Proceedings of the National...protein will be expressed in the same cell, but not necessarily at the same subcellular compartment as the...

  8. A Needle in a Base-Stack: Cas9 Structural Biology

    Type
    Blog Post
    ... it can rotate about 140 degrees between inactive and active conformations (Bravo et al., 2022; Sternberg...cutting efficiency and are important for robust activity in cells (Jiang & Doudna, 2017). Within an interior...non-target DNA strand is important for stabilizing the active conformation of the HNH domain, ensuring all binding...the HNH domain also allosterically controls RuvC activity through a hinge region between the two (Jiang ...RuvC can cleave DNA, which helps limit off-target activity. What next? It’s not actually clear! In vitro,...finish:   Figure 9: Cartoon summary of Cas9 activity. The apo enzyme is shown in the open conformation...a valuable laboratory tool are still an area of active research. New details are still being uncovered...

  9. CRISPR History and Development for Genome Engineering

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    Collection
    ... at specific loci. Activate or Repress : dCas9 fused to transcriptional activators or repressors can modulate...processed into individual crRNAs by a special trans-activating crRNA (tracrRNA) with homology to the short palindromic...for gene knockout, as well as transcriptional activation or repression. Combined with the power of next-generation...gRNAs: Truncated gRNAs display less off-target activity. High fidelity Cas9s: Three high fidelity Cas9...eSpCas9, and HypaCas9 display very low off-target activity due to rationally designed mutations. Researchers...CRISPR-Mediated Control of Gene Repression and Activation. Cell . 159(3):647-6. PMID: 25307932 Gilbert ...targeting multiple loci can be easily expressed in the same cell or organism. The ability to precisely target...

  10. 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
    ... on bacterial expression systems here) and the activity of the plant compounds on these proteins will ... be tested. Investigating changes in neuronal activity after ischemic brain injury Dr. Gohar Tsakanova...image cortical neurons and study changes in neural activity after ischemic brain injury. Additionally, optogenetic...construct plasmids containing promoter regions that activate fluorescent protein transcription in response ...robot that will “swim” in the river, take water samples at specific locations, measure fluorescent signals...

  11. Hot Biosensors 2022: Year-End Roundup

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    Blog Post
    ... A, et al. Glutamate indicators with improved activation kinetics and localization for imaging synaptic...signaling receptors, but the options for measuring activity of specific RTKs are limited. In a new preprint... a modular system for monitoring specific RTK activation in vivo (Farahani et al. 2022). The authors take...phosphorylated state of an immunoreceptor tyrosinase-based activation motif (ITAM). By fusing ITAM to an RTK of interest...localizes to the plasma membrane only when the RTK is activated and the ITAM phosphorylated. To demonstrate that...pYtag fluorescent biosensors for monitoring RTK activation by shift in cellular localization. Image adapted...to the range of concentrations expected in your samples. Higher affinity isn’t always better, especially...

  12. Fluorescent Proteins 101: GFP Fusion Proteins - Making the Right Connection

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    Blog Post
    ...other proteins, it may still block the function or activity of the tagged protein because of its size. This...Determining other aspects of functionality (catalytic activity or interactions) can be challenging. If a fusion... visualization of heterotrimeric G protein Gq activation in living cells." BMC biology 9.1 (2011): 32.... Jin, and Peter Devreotes. "Receptor-mediated activation of heterotrimeric G-proteins in living cells....al. "A FRET-based biosensor for measuring Gα13 activation in single cells." PloS one 13.3 (2018): e0193705...for robust measurement of Gαi1, Gαi2 and Gαi3 activation kinetics in single cells." PloS one 11.1 (2016...conclusive evidence that the fusion operates in the same way as the native protein. Although a fluorescent...

  13. Plasmids 101: Biotinylation

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    Blog Post
    ... is a cofactor that in its active form is covalently attached to the active site of key metabolic enzymes...is covalently bonded to a specific lysine at the active site of a newly synthesized enzyme. Biotin protein...polypeptides to a substrate Flow cytometry/fluorescence-activated cell sorting (FACS) In situ hybridization Methods...2388593 7. Holden, M. A., & Cremer, P. S. “Light activated patterning of dye-labeled molecules on surfaces...Wang, Y.-H., Lee, C.-Y., & Li, P.-C. “Buoyancy-Activated Cell Sorting Using Targeted Biotinylated Albumin...side chain that can be easily manipulated to make reactive species without affecting the avidin binding site...pre-incubated with free avidin can be added to your sample. Any free biotin-binding sites on this complex ...

  14. Hot Plasmids: Winter 2025

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    Blog Post
    ...Electrical and optical recordings of spontaneous activity in human stem cell-derived neurons expressing ...used in different illumination regimes with fast activation kinetics and high responsivity near the resting...Luciferase-fused detector that specifically binds to active (GTP-bound) Gα. Gα-YFP assembles into functional...heterotrimers with endogenous Gβγ, enabling detection of activation triggered by endogenous GPCR. Example applications...Direct interrogation of context-dependent GPCR activity with a universal biosensor platform. Cell, 187...recombinant antibody in human postmortem brain samples. Results show positive staining in tissues from...

  15. 27 Hot Plasmids from 2016

    Type
    Blog Post
    ... of robust CRISPR activators We previously reported on a novel Cas9-based activator developed by the Church...facilitates robust activation of target genes (20-40x greater than standard dCas9-VP64 activators) when introduced...Although the VPR activation domain is an incredible tool for researchers trying to activate target genes,..., which then recruit an MS2-fused hyperactive variant of activation-induced cytidine deaminase (AID). ...(Q3) | December 2016 (Q3) MARCH 2016Enhancer activity mapping with STARR-seq   Although many technologies... identification and quantification of enhancer activity have remained challenging due to the variable ...alternative method, called STARR-seq (self-transcribing active regulatory region sequencing) that tests enhancer...

  16. CRISPR 101: RNA Editing with Cas13

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    Blog Post
    ...knockdown, it still displays substantial off-target activity. By mutating residues in ADAR2 that interact with...the RNA substrate resulted in 15% C-to-U editing activity. They then performed sixteen rounds of directed...ADAR2DD to further optimize cytidine deaminase activity. While RESCUE can perform C-to-U edits, the authors...editing efficiency. They note that RESCUE is most active in a 30 nt guide when C or U base-flips are present...East-Seletsky, Alexandra, et al. "Two distinct RNase activities of CRISPR-C2c2 enable guide-RNA processing and...editing. To increase A->G editing, they employed hyperactive ADAR constructs, like ADAR2DD(E488Q). They also...deaminate cytidine and used dRanCas13b (catalytically inactive Cas13) to target the cytosine deaminase. Three...

  17. Bacterial Toxin-antitoxin Systems as Molecular Biology Tools

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    Blog Post
    ...processes and an antitoxin that suppresses the toxin’s activity. The key to these systems is that the toxin is...death. Conversely, CcdA binds to and blocks the activity of CcdB. Applications of ccdB in plasmid cloning...continual production of MazE, the toxin becomes active. Shen Yang’s lab made use of mazF as a counterselectable...systems consist of two enzymes that have opposing activities, it is not surprising that they also play toxin-antitoxin...toxin-antitoxin system, they were able to monitor lacZ activity over 48 hours in vitro without antibiotic selection... introduce multiple genetic modifications in the same strain. MazF expression is under control of the ...

  18. Top Requested Lentivirus and AAV of 2016

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    Blog Post
    ...These can be used to investigate the effects of activating particular signaling pathways in specific cell... et al., 2003), which can then be exclusively activated by the small molecule clozapine N-oxide (CNO) ...regions of mice, rats, or non-human primates. To activate the DREADD, CNO is delivered either through drinking...Maratos-Flier E, Roth BL, Lowell BB. Rapid, reversible activation of AgRP neurons drives feeding behavior in mice...systems typically express Cas9 and a gRNA from the same plasmid. LentiCas9-blast virus can be used to generate...

  19. Searchable and Sortable gRNAs for Your Next CRISPR Experiment

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    Blog Post
    ...gRNAs that were used for activation in a human system, search for ‘sapiens activate’ to filter the table ...applications we currently have data for are: cut, activate, interfere, visualize, nick, purify, tag, scaffold... consider testing multiple gRNAs targeted to the same locus to ensure their effects are specific to that...

  20. Deep Mutational Scanning with One Pot Saturation Mutagenesis

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    Blog Post
    ...phage display assays, ubiquitination activity assays, toxin activity, small molecule binding, growth rate...discover unexpected epistatic changes in protein activity. See Natarajan et al for an example of this. .... Stephany, and Stanley Fields. “Measuring the Activity of Protein Variants on a Large Scale Using Deep...These enzymes, Nt.BbvCI and Nb.BbvCI, recognize the same restriction site, but nick opposite strands of DNA...: The second mutant strand is synthesized in the same manner as the first mutant strand, but a universal...followed by the top strand. Both produced about the same number of positive clones. See Supplementary Note...primers? The key is to design primers that are the same sense as the strand of DNA degraded in the first...
Showing: 81 - 100 of 356 results