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We narrowed to 356 results for: SAM activators

Showing: 21 - 40 of 356 results

  1. 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...

  2. 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...

  3. 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...

  4. 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,...

  5. CRISPR Plasmids - Bacteria

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    Collection
    ... PI Publication Activate Catalytically dead dCas9 fused to a transcriptional activator peptide can increase...Edit Nick Prime Edit Transcriptional Regulation Activate Interfere Epigenetics RNA Targeting RNA Targeting...Design your gRNA sequence to direct the dCas9-activator to promoter or regulatory regions of your gene...separate gRNA expression plasmid to target the dCas9-activator to your specific locus. ID Plasmid Gene/Insert...target RNA sequence, they adopt an enzymatically active state and can bind and cleave additional RNAs regardless... regardless of homology to the crRNA. This activity provides a stark contrast to Cas9 and Cpf1, which ...cleaved. This non-specific cleavage is thought to activate programmed cell death or dormancy for phage-infected...

  6. 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...

  7. 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...

  8. CRISPR Plasmids - C. elegans

    Type
    Collection
    ... PI Publication Activate Catalytically dead dCas9 fused to a transcriptional activator peptide can increase...Edit Nick Prime Edit Transcriptional Regulation Activate Interfere Epigenetics RNA Targeting RNA Targeting...Design your gRNA sequence to direct the dCas9-activator to promoter or regulatory regions of your gene...separate gRNA expression plasmid to target the dCas9-activator to your specific locus. ID Plasmid Gene/Insert...protein and a target-specific gRNA in the same cell at the same time. Single plasmids containing both the...

  9. 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 ...

  10. CRISPR Plasmids - Empty gRNA Vectors

    Type
    Collection
    ... BbsI yes, activate, N-terminal S. pyogenes Zhang PX856 62888 Mammalian BbsI yes, activate, C-terminal...Bacteria BsaI yes, activate S. pyogenes Marraffini pWJ68 46571 Bacteria BsaI yes, activate S. pyogenes Marraffini...yes, activate S. pyogenes Luikart pRubiG-T2A-Cas9 75348 Mammalian/Retroviral from pXL yes, activate S. ... none S. pyogenes Boxem lentiSAMv2 75112 Mammalian/Lentiviral U6 yes, activate S. pyogenes Blast Zhang...Edit Nick Prime Edit Transcriptional Regulation Activate Interfere Epigenetics RNA Targeting RNA Targeting...-in-one vector, but their function (cut, nick, activate, interfere, etc.) is limited to that of the Cas...Cas9. We have plasmids with Cas9 that can cut, activate, interfere, or nick. Selection , such as Puromycin...

  11. 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...

  12. 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...

  13. CRISPR Plasmids - Plants

    Type
    Collection
    ... PI Publication Activate Catalytically dead dCas9 fused to a transcriptional activator peptide can increase...Edit Nick Prime Edit Transcriptional Regulation Activate Interfere Epigenetics RNA Targeting RNA Targeting...Design your gRNA sequence to direct the dCas9-activator to promoter or regulatory regions of your gene...separate gRNA expression plasmid to target the dCas9-activator to your specific locus. ID Plasmid Gene/Insert...S. pyogenes Yang 50579 pBUN6A11 OsU3 BsaI yes, activate S. pyogenes Bar Chen 50580 pBUN6I11 OsU3 BsaI ...pyogenes Bar Chen 50586 pHSN6A01 AtU6-26 BsaI yes, activate S. pyogenes Hyg Chen 50587 pHSN6I01 AtU6-26 BsaI...protein and a target-specific gRNA in the same cell at the same time. Single plasmids containing both the...

  14. CRISPR Plasmids - Drosophila

    Type
    Collection
    ... PI Publication Activate Catalytically dead dCas9 fused to a transcriptional activator peptide can increase...Edit Nick Prime Edit Transcriptional Regulation Activate Interfere Epigenetics RNA Targeting RNA Targeting...Design your gRNA sequence to direct the dCas9-activator to promoter or regulatory regions of your gene...separate gRNA expression plasmid to target the dCas9-activator to your specific locus. ID Plasmid Gene/Insert...protein and a target-specific gRNA in the same cell at the same time. Single plasmids containing both the...

  15. 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...

  16. CRISPR Plasmids - Yeast

    Type
    Collection
    ... PI Publication Activate Catalytically dead dCas9 fused to a transcriptional activator peptide can increase...Edit Nick Prime Edit Transcriptional Regulation Activate Interfere Epigenetics RNA Targeting RNA Targeting...Design your gRNA sequence to direct the dCas9-activator to promoter or regulatory regions of your gene...separate gRNA expression plasmid to target the dCas9-activator to your specific locus. ID Plasmid Gene/Insert...protein and a target-specific gRNA in the same cell at the same time. Single plasmids containing both the...Selectable Marker PI Publication Purify A catalytically inactive Cas9 (dCas9) can be used to purify a region of...

  17. 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...

  18. Fluorescent Protein Guide: Biosensors

    Type
    Collection
    ...signal the presence of specific biomolecules, the activity of specific genes and cellular processes, or other...) High-performance calcium sensors for imaging activity in neuronal populations and microcompartments....GCaMP6f/m/s calcium sensors for imaging neural activity (Constitutive or Cre-dependent) Ultrasensitive...Ultrasensitive fluorescent proteins for imaging neuronal activity. Nature. 2013 Jul 18;499(7458):295-300. Douglas ...expressing GCaMP6 and mRuby2 for imaging neural activity (GCaMP6f/m/s) (Constitutive or Cre-dependent) ...Optimization of a GCaMP calcium indicator for neural activity imaging. J Neurosci. 2012 Oct 3;32(40):13819-40...genetically encoded calcium sensor Imaging neural activity in worms, flies and mice with improved GCaMP calcium...

  19. Bacterial Expression Systems

    Type
    Collection
    ...particular protein can activate expression from that element. When this activation requires a transcription...Expression Species PI 14474 pRU1161 Promoter activity GUS activity and fluorescence (mRFP1) Gram-negative bacteria...bacteria Philip Poole 14473 pRU1156 Promoter activity GUS activity and fluorescence (GFPmut3.1) Gram-negative...Promoter activity Fluorescence (GFP+) Gram-negative bacteria Philip Poole 14460 pOT2 Promoter activity Fluorescence...Promoter activity Fluorescence (mCherry) Mycobacterium sp. Tanya Parish 24657 pASTA3 Promoter activity Fluorescence...Purification To study the function, structure, and activity of a protein it is generally necessary to purify...engineered strain for a B2H system used to screen the activity of arrays of zinc finger nucleases. Addgene Blog...

  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 356 results