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We narrowed to 347 results for: SAM activators

Showing: 21 - 40 of 347 results

  1. Brzezinski Lab CRISPR Collection

    Type
    Collection
    .../Cas9 based approach to knockout, repress, and activate gene targets. To accomplish this they have taken...sequence repression dCas9-VPR fusion for gene activation CRISPR/Cas9 Plasmid Variants Figure 1: Schematic...Plasmids 175571 and 175572 express dCas9-VPR for activation. Usage Guide sequences can be designed using ... a unique enhancer and either Ascl1 or Neurog2 activity . Development, 148(12). doi: 10.1242/dev.199399...sequence: Up to two guides can be expressed from the same plasmid by utilizing one of the dual guide shuttle...

  2. Tetracycline Inducible Expression

    Type
    Collection
    ...tetracycline-controlled transactivator (fusion of TetR with VP16 transcriptional activation domain); rtTA: reverse...tetracycline-controlled transactivator (fusion of mutated TetR with VP16 transcriptional activation domain). For ...tetracycline-controlled transactivator (tTA) was created by fusing TetR with the activation domain of the herpes...stronger expression of the downstream gene via its activation domain than simply removing a repressor. Tet-... to doxycycline and three copies of a minimal activation domain from VP16 for increased induction of gene...further improved rtTA variants with lower background activity and greater sensitivity to doxycycline (Zhou et...interest. If your gene of interest should be generally active and only turned off occasionally, use Tet-Off: ...

  3. Antibody Guide

    Type
    Collection
    ...fluorophores are available in a range of colors activated by different wavelengths, allowing for easy differentiation...conjugated with HRP. Wash and block the membrane. Activate the HRP with a substrate kit. Image the membrane...antibody, followed by incubation and wash steps. Activate the conjugate and read the output via a plate ... antigen and the conjugate, typically HRP, is activated and its output measured. This method can be performed...secondary antibody is added. The conjugate is activated and the output is read. A standard curve with ...deposited into tubes or plate wells) by fluorescence activated cell sorting (FACS). Figure 7: Flow cytometry ...application Collect and lyse samples to make proteins available. Tissue samples may need additional processing...

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

    Type
    Blog Post
    ...other hand, recruits transcriptional activators such as VP64, SAM, or SunTag to specific ncRNA loci to...DNA, CRISPR interference (CRISPRi) and CRISPR activation (CRISPRa) are powerful tools. CRISPRi uses a ...insertions or deletions in the DNA sequence. This same approach is effective to target short ncRNAs, such...useful for targeting thousands of long ncRNAs at the same time (Liu et al. 2018). However, removing large ...

  5. 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...

  6. CRISPR History and Development for Genome Engineering

    Type
    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...

  7. Lighting Up Cell Signaling with Photoswitchable Kinases

    Type
    Blog Post
    ...and pa= photoactivation) domains on either side of the active site of a constitutively active MEK1. This...localization serves as an indicator of MEK1 activity. When MEK1 is active, it phosphorylates the ERK KTR and red...and gut cells of C. elegans causes the same phenotype. Activation of the Raf-MEK-ERK signaling pathway ...Protein Activity with GFP Resources at Addgene.org Find KTR reporter plasmids Find photoactivatable fluorescent...signaling networks can also be non-specifically activated and, like the game of telephone, often the signal...intermediate messengers. Optical control of kinase activity can provide greater spatiotemporal resolution ...system, light is used to turn “on” and “off” MEK1 activity by attaching two dimerizing photoswitchable Dronpa...

  8. Viral Vectors 101: Chemogenetics

    Type
    Blog Post
    ...with less endogenous activation and reduced off target activation by ligands. PSAMs themselves are modified...engineered receptors that are only activated by otherwise inert drugs. Once activated, chemogenetic receptors hijack...First Structure–Activity Relationship Studies for Designer Receptors Exclusively Activated by Designer Drugs...DREADDs and PSAMs. Using DREADDs DREADDs DREADDS, or designer receptors exclusively activated by designer...provides a noninvasive method for manipulating cell activity, especially neuronal firing. What is chemogenetics...chemogenetics Because chemogenetic receptors are activated through administered drugs that must diffuse throughout... throughout the body, the onset of receptor activation is delayed and takes on the order of minutes. Studies...

  9. Chemogenetics vs. Optogenetics: Which Method Should I Choose?

    Type
    Blog Post
    ...delivery is the same in both cases, but activation depends on small molecule activation (chemogenetics)... Chemogenetics Timing Fast activation/deactivation Prolonged activation Targeting Further restricted...response or track the exact time of activation. Receptor activity is also drawn out as drugs can remain...offer the same level of control over timing. Because chemogenetics relies on chemical activation and the...neuroscience by granting targeted control over neuronal activity. While they serve similar purposes, both techniques...milliseconds, and can precisely link a neuron's activation to behaviors. Scientists continue to engineer...drugs throughout the body, the onset of receptor activation takes minutes. As a result, it is not possible...

  10. New Tools Enable CRISPRa for Neuroscience Applications

    Type
    Blog Post
    ...Suntag” Cas9 activator that recruits up to twenty copies of the p65-HSF1 transcriptional activator to each ...transcriptional activation of multiple genes in the brain using CRISPR–dCas9-activator transgenic mice...expression of the SPH activator paired with AAV sgRNA delivery make the SPH mouse an attractive tool to probe ...recent papers describe the development of CRISPR activation (CRISPRa) tools for neuroscience, including transgenic...neuroscience research. What is CRISPR-based activation? To summarize, CRISPRa uses a nuclease-dead variant...and engineered combinations of transcriptional activators to induce gene transcription at genomic sites... vitro settings due to the large size of Cas9 activators, which largely precludes delivery by adeno-associated...

  11. Viral Vectors 101: Optogenetic Tools

    Type
    Blog Post
    ... described the first light-activated tool for controlling neuronal activity, channelrhodopsin-2 (ChR2)...exposed to blue light, this channel activates neuronal activity by opening and allowing positively charged...cellular activity with exquisite precision using only light. Optogenetics, or light activated tools, can...either activate or inhibit a specific receptor or pathway.  Light-gated ion channels Light activated ion ...ChR2) activation by light showing ions flowing with the concentration gradient, and of activation inhibitory... to prevent activation of both tools with each light source used.   Channel and activation kinetics Some...ChR2), a blue light-activated cation channel, from the archaebacteria Chlamydomonas reinhardtii. When exposed...

  12. CRISPR 101: Anti-CRISPR Proteins Switch Off CRISPR-Cas Systems

    Type
    Blog Post
    ...way to shut off Cas’s activity. The concern is that the longer Cas remains active in a cell, the greater...switch and turn off CRISPR activity. The ability to turn off CRISPR activity is an important step towards...Cas9 activity in HEK293 by reducing editing from ~30% in control samples to 0-10% in Acr samples. Acr ... Cas proteins can be used for genome editing, activating gene expression, repressing gene expression, ...edits to occur. Although methods to switch on Cas activity using light or drugs have been developed, the ...proteins are highly diverse but most block CRISPR activity one of three ways: Inhibit DNA binding Example... Anti-CRISPR proteins (Acr's) can block CRISPR activity in many ways such as inhibiting crRNA loading,...

  13. Tips for Screening with Yeast Two Hybrid Systems

    Type
    Blog Post
    ...transcriptional activators: that the DNA binding domain (DBD) and transcriptional activation (TA) domains...discovery in the field of eukaryotic transcriptional activation and cell biology. During this period, proteins...expressed as fusions that retain their individual activities (1). Researchers also discovered the modular ...domains retain their individual activities when separated (2), and that DBD and TAs from different systems ...protein shown is still able to bind to the upstream activation sequence (UAS) of its target gene, in this case...of whether it interacts with a transcriptional activator (1a) or a repressor (1b). Scientists took advantage...commonly referred to as the “bait”, Figure 2b) cannot activate reporter gene expression on its own, but, if the...

  14. Plasmids 101: Repressible Promoters

    Type
    Blog Post
    ... are always active and thus referred to as constitutive promoters. Others are only active under specific...Repressible: An activator protein is bound, turning transcription ON. When a repressor binds the activator protein...concentrations are low, pADH1 displays low promoter activity (~20% activity of the strong yeast promoter pTEF). As ...or when that promoter is active. In this way, you can interrogate the activity of uncharacterized promoters...or VP16 activation domain results in chimeric GAL80-repressible and GAL80-insensitive activators of lexAop...introduced the second generation QF2 activators. These activators solve the problem of broad QF expression-induced...tetracycline-controlled transactivator (tTA). They fused TetR with the transcriptional activation domain VP16 taken...

  15. Plasmids 101: Inducible Promoters

    Type
    Blog Post
    ...promoter is inactive because the activator protein cannot bind. After an inducer binds to the activator protein...direct activation. In this system, the activator rtTA (reverse tetracycline-controlled transactivator) is... are always active and thus referred to as constitutive promoters. Others are only active under specific...OFF state, the promoter is inactive because a bound repressor protein actively prevents transcription. Once...protein, the activator protein can bind to the promoter, turning it ON and initiating transcription.     ...derivatives serve as inducing agents to allow promoter activation. One of the most commonly used prokaryotic promoters...repressor (lacI protein) for transcription to be activated. In the presence of lactose or lactose analog ...

  16. Optogenetics + CRISPR, Using Light to Control Genome Editing

    Type
    Blog Post
    ... optically active saCas9 protein. Stop light! Photoactive anti-CRISPRs halt Cas9 activity Anti-CRISPR...proteins heterodimerize and move the activator into position to activate gene transcription. The researchers...light-dependent activity to transcription, they fused BldD to the transcriptional activators p65 and VP64...can interfere with activity. The resulting system was coined as CRISPR–Cas9 activity switching via a novel...generate a photoactivatable nickase and a photoactivatable repressor (dCas9). The activity of all variants...Shining light on transcriptional activation using dCas9 Initial photoactivatable CRISPR systems published in...genomic anchor - an inactive, dead Cas9 protein (dCas9) fused to CIB1; and 2) the activator - the CRY2 photolyase...

  17. 28 Hot Plasmid Technologies from 2015

    Type
    Blog Post
    ...approach can also be used for activation of specific genes using dCas9-VP64 activator fragments (dCas9(C)-FKBP...endogenous gene activation. This system works by bringing together an anchor protein and activator protein to...portion of CRY2) fused to the p65 activation domain, which activates transcription of downstream genes...the targeted activation of endogenous genes and detail the time course of gene activation using this system...2015 Feb 23. A novel dCas9 activator capable of unprecedented activation of target genes A group of researchers...fused to a VP64 activator domain and targeted toward endogenous promoters to activate transcription. Chavez...synergistic effect on target gene activation. The novel dCas9-VPR activator is capable of enhancing target...

  18. How to Design Your gRNA for CRISPR Genome Editing

    Type
    Blog Post
    ...balance maximizing on-target activity while minimizing off-target activity, which sounds obvious but can...to use a less-active gRNA that targets a truly unique site in the genome, or a more-active gRNA with one...that your gRNA sequence is designed to be highly active and reduce off targets. For CRISPRa and CRISPRi...to generate the desired edit. The wrench: Gene activation and inhibition by CRISPRa and CRISPRi Finally...expression at the level of transcription – CRISPRa (activation) and CRISPRi (inhibition) technologies – a nuclease-dead...100nt window downstream of the TSS gives the most activity. Thus, a given gene will only have a dozen or ...features to nominate gRNAs that are likely to be active, not only for SpCas9 but also for some other Cas...

  19. SARS-CoV-2/COVID-19 Detection Methods Based on CRISPR/Cas

    Type
    Blog Post
    ...sequences, Cas12a activity gets activated. This unleashes indiscriminate ssDNA cleavage activity which cleaves...sequence to bind to the sample and upon binding, gets activated. This activation results in a trans cleavage...detection.  Highlights: Cas13a is not catalytically active in the presence of two or more mismatches in the...complementary region in the target sequence and this activity results in the trans cleavage of the ssDNA-FQ ... given fluorescently labelled substrate, gets activated and performs non-specific collateral cleavage....cleavage by the enzyme and instead relies on its activity on the target regions thus making it an affinity-based...the Sashital Lab, Iowa State University. She is actively interested in science communication and enabling...

  20. Choosing a CRISPR Nuclease: Site Accessibility, Specificity, and Sensitivity

    Type
    Blog Post
    ...by calculating the on-target activity divided by the off-target activity. Figure 2: Specificity...measure of off-target activity (left) while sensitivity is the measure of on-target activity (right). Cas9...Cas9 exhibits high on-target activity, but also high off-target activity, which may be undesirable in ...Specificity is the assessment of the off-target activity of Cas9. This can occur when Cas9 cleaves a site...specificity. Sensitivity is a measure of the on-target activity of the Cas9 nuclease. This can be determined by...Both of the enzymes showed decreased off-target activity relative to Cas9. Since then, the Joung and Jennifer...nuclease showed decreased or similar off-target activity, relative to eSpCas9 and SpCas9-HF across a number...
Showing: 21 - 40 of 347 results