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We narrowed to 642 results for: ato

Showing: 61 - 90 of 642 results

  1. Sonic Hedgehog? Sleeping Beauty? Learn About the Genes Behind Addgene’s Conference Room Names

    Type
    Blog Post
    ...Cancer Institute! Sonic Hedgehog, an important regulator of embryonic development The largest conference...the hedgehog signalling pathway, which is a key regulator of all bilateral animal development. SHH plays...publication, Ivics et al 1997. Spaghetti Squash, a regulator of cellular contraction You may think that a gene...vessels). Spaghetti squash (Sqh) is the Drosophila Regulatory Light Chain (RMLC) of nonmuscle myosin II (Karess... Jordan and Karess, 1997), which contains the regulatory regions that are necessary for myosin activity...monster, a meiotic arrest gene required for spermatogenesis One of our smaller conference rooms is named... meiotic arrest gene that is necessary for spermatogenesis. Flies that lack cookie monster are sterile...

  2. Zhang Lab CRISPR Page

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    Collection
    ...Expresses dCAS9-VP64 activator with 2A GFP 61423 : Expresses the MS2-P65-HSF1 activator helper complex with...screening library CRISPR/Cas9 Synergistic Activation Mediator (SAM) is an engineered protein complex for the...: lentiviral vector encoding the MS2-P65-HSF1 activator helper complex with a 2A Hygro resistance marker...JS, Konermann S, Trevino AE, Scott DA, Inoue A, Matoba S, Zhang Y, Zhang F. Cell . 2013 Sep 12;154(6):...

  3. Hot Biosensors 2022: Year-End Roundup

    Type
    Blog Post
    ...doi: 10.1038/s41592-022-01597-x. New glutamate indicator iGluSnFR3 Improving on the glutamate sensor iGluSnFR...PDGFR.codonopt    Aggarwal A, et al. Glutamate indicators with improved activation kinetics and localization...have developed a new genetically-encoded voltage indicator (GEVI) called JEDI-2P (Liu et al. 2022) optimized...Sustained deep-tissue voltage recording using a fast indicator evolved for two-photon microscopy. Cell. 2022 ...has generated a new genetically encoded calcium indicator using small monomeric NIR fluorescent proteins... of a Small Near-Infrared Fluorescent Calcium Indicator. Front Cell Dev Biol. 2022 Jun 29;10:880107. doi...they developed a family of fluorescent potassium indicators with a range of binding affinities (Torres Cabán...

  4. CRISPR Plasmids - C. elegans

    Type
    Collection
    ...gRNA sequence to direct the dCas9-activator to promoter or regulatory regions of your gene of interest....Catalytically dead dCas9 fused to a transcriptional activator peptide can increase transcription of a specific...separate gRNA expression plasmid to target the dCas9-activator to your specific locus. ID Plasmid Gene/Insert...

  5. CRISPR Meets Synthetic Biology: A Conversation with MIT’s Christopher Voigt

    Type
    Blog Post
    ...genes. We’ve created enough regulatory elements, CRISPRs, repressors, activators. We’ve been putting together...CRISPR/Cas genetic circuits are linked to the regulatory networks within E. coli host cells. ...in building genetic circuits is getting enough regulatory proteins that don’t interfere with each other...programmable that you can make a very large number of regulators. That basically means larger genetic circuits...that could conceivably be as large as natural regulatory networks. Addgene: What kinds of things could... pattern, shape. All of that is controlled by regulatory networks in the plant. If we want to access that...CRISPR is how much easier it is to work with than regulatory proteins. There are still things we haven’t solved...

  6. CRISPR Pooled gRNA Libraries

    Type
    Collection
    ...3rd 10 187,535 Advanced Catalogue of Epigenetic Regulators (ACER) 226117 Knockout Human Lu 3rd 10 8,205 ...Knockout Mouse Tolar 3rd ~4 78,637 Mouse Chromatin Regulator Library 200011 Knockout Mouse Griffin, Manguso...4 17,032 arrays Mouse Cardiac Transcriptional Regulators CRISPR Knockout Library 138015 Knockout Mouse...CRASP-Seq Gene KO Library 232069 Knockout Human Gonatopoulos-Pournatzis 3rd Varies 95,893 CRASP-Seq BE Tiling...Tiling Library 232070 Base Editing Human Gonatopoulos-Pournatzis 3rd Varies 8,594 CROPseq-iT7 Human CRISPR...

  7. Nuclear Receptor Signaling Atlas (NURSA) Plasmid Guide: Nuclear Receptors

    Type
    Collection
    ...HNF4alpha, MODY, MODY1, NR2A1, NR2A21, TCF, TCF14 hepatocyte nuclear factor 4, alpha NR0B1 Plasmids NR0B1,....5, NR1C1, PPAR, PPARalpha, hPPAR peroxisome proliferator-activated receptor alpha PPARD Plasmids PPARD..., NR1C2, NUC1, NUCI, NUCII, PPARB peroxisome proliferator-activated receptor delta PPARG Plasmids PPARG... NR1C3, PPARG1, PPARG2, PPARgamma peroxisome proliferator-activated receptor gamma RARA Plasmids RARA,...

  8. CRISPR Plasmids - Drosophila

    Type
    Collection
    ...gRNA sequence to direct the dCas9-activator to promoter or regulatory regions of your gene of interest....Catalytically dead dCas9 fused to a transcriptional activator peptide can increase transcription of a specific...separate gRNA expression plasmid to target the dCas9-activator to your specific locus. ID Plasmid Gene/Insert...

  9. CRISPR Plasmids - Yeast

    Type
    Collection
    ...gRNA sequence to direct the dCas9-activator to promoter or regulatory regions of your gene of interest....Catalytically dead dCas9 fused to a transcriptional activator peptide can increase transcription of a specific...separate gRNA expression plasmid to target the dCas9-activator to your specific locus. ID Plasmid Gene/Insert...

  10. Early Career Researcher Toolbox: Free Online Molecular Biology Tools

    Type
    Blog Post
    ...In-Fusion cloning molar ratio calculators Lab math calculators These calculators can help you with common...lab. Molarity calculator Promega's Biomath Calculators Primer resuspension calculator (sign-in required...Golden Gate Reactions Cloning ligation reaction calculators These links can help you calculate how much insert...assembly, or In-Fusion cloning.  NEB Ligation Calculator: This tool works for any cloning method. You ...required.)  DailyCalcs Science Calculator app for iPhone Reagent and Data Repositories These links can help...

  11. AAVED

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    Collection
    ... Institute Katherine Matho Cold Spring Harbor Laboratory Gowan Tervo Janelia Research Campus Sarada Viswanathan...Regulation Ben Deverman 2:00 PM Cre-DOG and Combinatorial Strategies Connie Cepko 2:45 PM Break 3:00 PM...In vivo data: Retrograde hSyn1-GCaMP6f-P2A-nls-dTomato Detailed protocol: AAV Production in HEK293 Cells...Institute Katherine Matho, Cold Spring Harbor Laboratory Gowan Tervo and Sarada Viswanathan, Janelia Research...

  12. Rewiring Metabolic Circuitry with CRISPR RNA Scaffolds [Video]

    Type
    Blog Post
    ...transcriptional regulator to modulate expression at that locus. The use of regulators allows a greater...shifted in recent years toward tinkering with regulatory networks and multiplex gene control. However,...approaches are often met with the challenges of regulatory bottlenecks and their scope is limited by the...scaffolds that can recruit various transcriptional regulators (Zalatan et al. 2015).  These scaffolds are stretches...levels dependent upon the type of transcriptional regulator or the number of designed scaffold binding sites...with distinct programs controlled by each dCas9 regulatory switch. By no means limited to the field of metabolic...scaffolds can recruit various transcriptional regulators to alter gene expression at the targeted site...

  13. CRISPR Plasmids - Bacteria

    Type
    Collection
    ...gRNA sequence to direct the dCas9-activator to promoter or regulatory regions of your gene of interest....Catalytically dead dCas9 fused to a transcriptional activator peptide can increase transcription of a specific...separate gRNA expression plasmid to target the dCas9-activator to your specific locus. ID Plasmid Gene/Insert...

  14. Addgene's Cancer Collection Pages Connect the Research Community

    Type
    Blog Post
    ...to their laboratory. A single mouse click on a gene of interest will direct the investigator to a list... cancer are yielding promising results in the laboratory, and Addgene is accelerating this research by...proto-oncogenes and tumor suppressors to the accelerator and brake pedal of a vehicle, respectively. In... (now an oncogene), this is analogous to the accelerator of a vehicle being stuck down. Similarly, a mutation...cells still need to successfully infiltrate the circulatory or lymph systems to spread, or metastasize. Many...Many cancerous cells will die trying, as the circulatory and lymph systems are not ideal environments ...involved in the Ras pathway alone and thousands of investigators studying them, how do we ensure that the research...

  15. CRISPR Plasmids - Mammalian Expression

    Type
    Collection
    ...gRNA sequence to direct the dCas9-activator to promoter or regulatory regions of your gene of interest....Catalytically dead dCas9 fused to a transcriptional activator peptide can increase transcription of a specific...separate gRNA expression plasmid to target the dCas9-activator to your specific locus. ID Plasmid Gene/Insert...

  16. CRISPR Plasmids - Plants

    Type
    Collection
    ...gRNA sequence to direct the dCas9-activator to promoter or regulatory regions of your gene of interest....Catalytically dead dCas9 fused to a transcriptional activator peptide can increase transcription of a specific...separate gRNA expression plasmid to target the dCas9-activator to your specific locus. ID Plasmid Gene/Insert...

  17. CRISPR 101: Which Cas9 Do I Choose for My CRISPR Experiment?

    Type
    Blog Post
    ...target genes using dCas9-based activators The simplest of the dCas9 activators consists of dCas9 fused to ... dCas9 and antibody-activator fusion proteins. The Synergistic Activation Mediator complex, generally ...exploited to localize factors such as transcriptional activators and repressors to target genes. Repressing target...activation domain - VP64.  Later generations of activators include the SunTag system which facilitates recruitment...generally the strongest activator, consists of a dCas9-VP64 fusion and a modified gRNA that is capable of interacting...with an additional RNA-binding transcriptional activator. Plasmids for activating target genes in a variety...fusion, both targeted to the promoter region. Cas9 activators include Cas9 fused to VP64, VP64 along with other...

  18. Hot Plasmids - October 2022

    Type
    Blog Post
    ... voltage indicator: JEDI-2P  by: Brian O'Neill  The François St-Pierre lab and collaborators have successfully...what you'll find in this post: Improved voltage indicator: JEDI-2P Novel class of light-gated potassium...successfully developed an improved voltage indicator (GEVI) called JEDI-2P. Using a custom, high-throughput...predecessor GEVIs (called ASAP 1 and 2), they found an indicator that is optimized for 2-photon imaging of voltages...screen for improved genetically-encoded voltage indicators (GEVIs) based on the ASAP sensors. The resultant...Sustained deep-tissue voltage recording using a fast indicator evolved for two-photon microscopy. Cell, 18, 3408... cations. B) Cortical slice of HcKCR1-EYFP and tdTomato expressed layer 2/3 neurons in mouse. C) Action...

  19. Recombinase-based State Machines Enable Order-dependent Logic in vivo

    Type
    Blog Post
    ...massive amount of information regarding which regulatory proteins, signaling molecules, mutations, and...paper (1). Lastly, it is possible to build gene regulatory RSMs (GRSMs, the subset of RSMs that regulate...also demonstrates that genetic parts (genes, terminators, and promoters) may be interweaved onto a GRSM...parts used in this paper (recognition sites, terminators, promoters, and genes) were built with flanking...them. We recommend using strong, directional terminators and promoters with buffer sequences on their ...ends (for insulation). A list of promoters and terminators used in our GRSM registers can be found in Table... parts (recognition sites, promoters, genes, terminators, etc) are composed. Fear not – the state machine...

  20. Plasmids 101: Inducible Promoters

    Type
    Blog Post
    ...the activator protein cannot bind. After an inducer binds to the activator protein, the activator protein...In this system, the activator rtTA (reverse tetracycline-controlled transactivator) is normally inactive...protein purification. When arabinose is absent, regulatory protein AraC binds O and I1 sites upstream of...addition to arabinose, cAMP complexed with cAMP activator protein (CAP) can also stimulate AraC binding ...research. Promoter Subtype Promoter Example Activator Addgene Plasmid/Kit Alcohol-regulated AlcA...

  21. Truncated gRNAs for Regulating Gene Expression

    Type
    Blog Post
    ...cut either strand of DNA. Traditional Cas9 regulators In addition to its utility in making targeted...to DNA, Cas9 can be reprogrammed to serve as a regulator of gene expression. Its catalytic domains can ...fused to effector domains such as transcriptional activators or repressors. These fusion proteins retain the... generate a potent Cas9-based transcriptional regulator without making any functional changes to Cas9....adapter capable of recruiting a transcriptional activator, robust regulation of gene expression is observed...the truncated gRNA recruits a transcriptional activator to the Cas9-gRNA complex. When this complex is...of the Addgene Scientific Advisory Board. His laboratory at Columbia University is focused advancing scientific...

  22. Transgenic Organisms, Cas9 Gene Drives, and Appropriate Safeguards

    Type
    Blog Post
    ...carrying a synthetic gene drive were to escape the laboratory, the drive could eventually spread through the...mistake would necessarily extend far beyond the laboratory and seriously damage public trust in scientists... can spread through populations of transgenic laboratory organisms but not wild organisms. For example...synthetic sequence will only spread in transgenic laboratory populations with that sequence2. Both methods...Reproductive confinement involves working with laboratory organisms that can't reproduce with wild ones...confinement seeks to keep the organisms in the laboratory. It varies by organism, but your local biosafety...population and immunizes it against the first drive. Laboratories interested in building candidate gene drives...

  23. Isolating Genomic Regions of Interest with the CRISPR System

    Type
    Blog Post
    ...the Tapscott Lab identified regulators of DUX4, a transcriptional regulator that when inappropriately expressed...activation using a fusion to a transcriptional activator such as VP64 (Gilbert et al., 2013), or repression...region to inhibit association of transcriptional activators (Qi et al., 2013). However, the fusion of dCas9...specific genomic region In 2013, the Hodaka Fujii Laboratory first described a method to purify a specific...lab is CRISPR affinity purification in situ of regulatory elements (CAPTURE) (Liu et al., 2017). This system...additional research conducted in the Hodaka Fujii Laboratory, please also see our interview with Dr. Fujii...immunoprecipitation identifies miR483 as an epigenetic modulator of IGF2 imprinting in tumors." Oncotarget 8.21...

  24. Simplify Cloning with in vivo Assembly

    Type
    Blog Post
    ...  Javier García-Nafría is an investigator scientist at the MRC Laboratory of Molecular Biology. He is ... Watson and Javier García-Nafría from the MRC Laboratory of Molecular Biology. Plasmid cloning is an essential...with successful recombination reported in all laboratory E. coli strains tested so far. The use of this...low cost and can be immediately adopted by any laboratory. All procedures are performed using the same...the homologous region (calculated using Oligo Calculator). Insertions - To insert short sequences of ... Watson and Javier García-Nafría from the MRC Laboratory of Molecular Biology. Jake Watson is a postdoctoral...postdoctoral researcher in neurophysiology at the MRC Laboratory of Molecular Biology. He is currently investigating...

  25. 6 Tips for Analyzing and Troubleshooting Sanger Sequencing Results

    Type
    Blog Post
    ...to always look closely at the trace file (or chromatogram) of the sequencing results you get back from...facility. When it comes to DNA sequencing the chromatogram is your visual control. And, like all controls...controls, missing out is a big mistake. Chromatograms come to the rescue for analyzing and troubleshooting...But look closer, the overlapping peaks in the chromatogram suggest the results are not as certain as the... of the following programs to view your .ab1 chromatogram file 4Peaks (Mac) SnapGene Viewer (Mac/PC) ...peak, don’t be shy. Feel free to edit it. Most chromatogram viewing programs (even the free ones) allow ...allowing us to share it with our readers! The chromatograms in this article were created with SnapGene. ...

  26. Hot Plasmids - August 2020

    Type
    Blog Post
    ...mDlx-GFP, hDlx-FLEX-GFP, hDlx-Flex-dTomato, mDlx-GCaMP6f, hDlx-GiDREADD-dTomato and mDlx-ChR2-mCherry. Find... a cell. The Goedhart lab, Gadella lab, and collaborators have an ongoing effort to improve genetically...by one-step immobilized metal ion adsorption chromatography (IMAC) using Ni-NTA magnetic beads. Workflow...multiplexed gRNAs and of dCas9 fusions. The dCas-KRAB activator and the dCas-VPR repressor are introduced into...an EGFP reporter. Expression of the dCas9-KRAB activator or dCas9-VPR repressor in hPSCs is also driven...expression. This includes a CRISPRi-based synthetic oscillator, bistable network, stripe pattern-forming incoherent...calcium sensors (jYCaMP1), variants of the calcium indicator jGCaMP7, that are optimized for excitation wavelengths...

  27. Plasmids 101: The Promoter Region – Let's Go!

    Type
    Blog Post
    ...controlled by still other regulatory factors: enhancers, boundary elements, insulators, and silencers; however...tetracycline operators. Transcription can be turned on or off depending on what tet transactivator is used....the promoter (RNA polymerase binding site) and operators (response elements). Promoters are about 100 to...expression Promoter from T7 bacteriophage plus lac operators Negligible basal expression when not induced. ...RNA polymerase, which is also controlled by lac operator. Can be induced by IPTG. Commonly found in pET...pET vectors. Very tightly regulated by the lac operators. Good for modulating gene expression through varied...

  28. COVID-19 Resources

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    Collection
    ...platforms to reliably detect nucleic acids at the atomolar level. For more information read Addgene's Blog...and open-access SARS-CoV-2 detection assay for laboratory and home testing. Kellner, et al. bioRxiv 2020.06.23.166397...Cathepsin L Functionally Cleaves the Severe Acute Respiratory Syndrome Coronavirus Class I Fusion Protein Upstream...TMPRSS2 augments entry driven by the severe acute respiratory syndrome coronavirus spike protein. . PCP4 - ...

  29. Tetbow: Bright Multicolor Labeling for Neuronal Tracing

    Type
    Blog Post
    ... introduced, stochasticity will result in a combinatorial expression of these three genes with different...inherently low. This is because the stochastic and combinatorial expression of fluorescent proteins is only possible...original method, we used the Tetracycline trans-activator system to boost the expression levels, and we ...we named the system Tetbow (Tetracycline trans-activator Brainbow) (Sakaguchi et al., 2018) (Figure 1)....sensory systems focusing on the functional and anatomical maturation of the olfactory system.      Takeshi...University. His lab has been studying functional and anatomical aspects of sensory circuit development using ...Livet, Jean, et al. "Transgenic strategies for combinatorial expression of fluorescent proteins in the nervous...

  30. Cultivating Community Science at BosLab

    Type
    Blog Post
    ...fully-equipped laboratory for citizen scientists. In addition to maintaining a laboratory to support both... of independent Do-it-Yourself (DIY) biology laboratories working towards enabling everyone to participate... set on top of filing cabinets, a rickety old incubator, a few pipettes, and some home-made equipment....Anyone can become a member of BosLab and previous laboratory experience is not required. Our members are skilled...have non-traditional scientists working in our laboratory! Don’t worry though, we make sure that everyone...a co-organizer at BosLab, a community biotech laboratory in Somerville MA. Follow her on twitter at @ZebraElement...
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