We narrowed to 903 results for: nes

...Li Norepinephrine nLight red and green GPCR-based NE indicators Sensitive multicolor indicators for monitoring...Norepinephrine GRAB_NE family of GPCR activation-based NE sensors A Genetically Encoded Fluorescent Sensor ...fluorescent biosensors to measure biomolecules or genes via FRET or other assays. Plasmid...specific biomolecules, the activity of specific genes and cellular processes, or other factors like environmental...Campbell Calcium Ratiometric calcium biosensors from nested reporters of green- and orange-emitting proteins...proteins Ratiometric Matryoshka biosensors from a nested cassette of green- and orange-emitting fluorescent...mScarlet-based calcium sensor with exceptional brightness, photostability, and multiplexing capabilities...

...Knockout Screening in Human Cells. Shalem O, Sanjana NE, Hartenian E, Shi X, Scott DA, Mikkelson T, Heckl...and easily manipulate specific genes. But what if you want to study genes all across the genome? Two new...screen for genes involved in bacterial toxin resistance, identifying 4 previously unknown genes.  “With ...library, which they call GeCKO, targets 18,080 human genes with 64,751 unique guide sequences to enable both...including Feng Zhang used the GeCKO library to identify genes essential for cell viability in cancer and pluripotent...In a melanoma model, they also went in search of genes involved in resistance to the cancer drug vemurafenib...guide RNAs (gRNAs) covering a total of 7,114 human genes. Unlike the GeCKO library, each library has been...

...expressing nLightG or nLightR before/after application of NE (10 μM) and corresponding pixel-wise ΔF/F0 heatmaps... expression of the indicators over white dashed lines. Scale bars, 10 μm (HEK293T), 20 μm (neurons). Figure...options for multiplexing in experiments, including ones that study alpha2-AR. Selectivity Norepinephrine...

...Feb;10(4):337-47. PubMed PMID: 12595892. 4. Sanjana NE, Shalem O, Zhang F. Improved vectors and genome-wide...virus can be used to generate Cas9-expressing cell lines, which can then be used both for screens (using ...

...Promoter pRSET OCaMP pRSET T7 pAAV-hSYN1-NES-OCaMP-WPRE pAAV hSyn pAAV-EF1a-FLEX(FLP)-...calcium, which is more than just the stuff that makes bones strong; it plays critical roles in nerve and muscle...power, but because they are non-tunable, their usefulness depends on selecting a fluorophore matched to...enabling two-photon imaging without compromising on brightness, ∆F/F0, or signal-to-noise ratio (Figure 3). ...signal change during calcium events), Greater brightness, Less signal distortion from photoswitching or...

...genome-wide libraries for CRISPR screening. Sanjana NE, Shalem O, Zhang F. Nature Methods. 2014 Aug;11(8.... After the monoclonal lines have been sufficiently expanded, screen the lines for transgene expression...transgene expression over time, as the lower expressing clones take over the polyclonal cell pool. Generating ...details, see our protocol for generating stable cell lines with lentivirus . Day 0: (optional) Seed cells for... cells Day 14–30: Analyze and expand monoclonal lines of interest Equipment Class II, Type A2 Biological...Heat-inactivated FBS 1X PBS pH 7.4 without calcium or magnesium, Corning 21-040-CV (cations can affect the attachment...pool: see our protocol for generating stable cell lines with lentivirus . Pro-Tip Because the polyclonal...

...Gootenberg JS, Abudayyeh OO, Platt RJ, Brigham MD, Sanjana NE, Zhang F (2017) Genome-scale CRISPR-Cas9 knockout...available reference sequence or plasmids containing genes with many modifications. We look for point mutations...

...specific genes using dCas9-VP64 activator fragments (dCas9(C)-FKBP-2xNLS-VP64 and dCas9(N)-FRB-NES). This...FKBP rapamycin binding domain of mTor (Cas9(N)-FRB-NES) resulting in a rapamycin-inducible Cas9 for genome...editing. Without rapamycin treatment, the Cas9(N)-FRB-NES fragment is actively shuttled out of the nucleus ...sequence. Treatment with rapamycin induces Cas9(N)-FRB-NES and Cas9(C)-FKBP-2xNLS dimerization and net influx...of downstream genes. Nihongaki et al describe the targeted activation of endogenous genes and detail the... promoter-less Gateway® entry clones to be used with other entry clones encoding the ORFs of interest:...neuron-like” phenotype when targeting genes involved in neurogenesis, which had proven challenging using...

...beta-hydroxylase (DBH, magenta) to label norepinephrine (NE) expressing neurons and GFP (green) to label neurons...’m willing to bet the answer would be along the lines of “we have a lot”. In fact, scientists often cite..., C. M., Leib, R. D., Cirolia, G., Thomas, D., Stamnes, S., Holt, K., Sinn, P., May, A. P., & Paulk, N...availability upon request differ across scientific disciplines. Scientific Data, 8(1), 1–11. https://doi.org...

...Gootenberg JS, Abudayyeh OO, Platt RJ, Brigham MD, Sanjana NE, Zhang F. Genome-scale CRISPR-Cas9 knockout and transcriptional...

...genome-wide libraries for CRISPR screening. Sanjana NE, Shalem O, Zhang F. Nat Methods. 2014 Aug;11(8):783...2nd-generation lentiviral system (Figure 2). The HIV genes that do remain are very important for viral production...make stable transgene-expressing or knockout cell lines. Many CRISPR plasmids are designed for lentiviral...integrate into the genome, they could promote oncogenesis by altering local gene expression. Self-inactivating...LTR that prevents aberrant activation of nearby genes. These safer vectors have become standard in gene...

...repress, visualize, and isolate genes. Activation or Repression of Target Genes Targeting dCas9 to transcription... and a user-defined ∼20-nucleotide spacer that defines the genomic target to be modified. You can therefore...CRISPR was originally employed to knock out target genes in various cell types and organisms, but modifications... ability to selectively activate/repress target genes, purify specific regions of DNA, image DNA in live...Multiplexing applications include editing multiple genes at once; using dual nickases to generate a knockout... used to knock out, activate, or repress target genes when paired with the appropriate Cas enzyme. Specific...CRISPR specificity. SpCas9 (from Streptococcus pyogenes ) is the most popular Cas endonuclease, with many...

...AAV-EF1a-BbTagBY Control Joshua Sanes AV-9-PV2454 45186-AAV9 AAV-EF1a-BbChT Control Joshua Sanes AV-9-PV2629 100043...Looger, Eric Schreiter AV-1-PV3844 100855-AAV1 pAAV.CAG.Flex.NES-jRGECO1b.WPRE.SV40 Biosensor GENIE, Douglas...Douglas Kim AV-1-PV3845 100856-AAV1 pAAV.Syn.Flex.NES-jRGECO1b.WPRE.SV40 Biosensor GENIE, Douglas Kim AV-1...-1-PV3846 100857-AAV1 pAAV.Syn.NES-jRGECO1b.WPRE.SV40 Biosensor GENIE, Douglas Kim AV-1-PV3847 100852-...100852-AAV1 pAAV.CAG.Flex.NES-jRGECO1a.WPRE.SV40 Biosensor GENIE, Douglas Kim AV-1-PV3848 100853-AAV1 pAAV....Biosensor GENIE, Douglas Kim AV-1-PV3849 100854-AAV1 pAAV.Syn.NES-jRGECO1a.WPRE.SV40 Biosensor GENIE, Douglas ...Douglas Kim AV-1-PV3850 100849-AAV1 pAAV.CAG.Flex.NES-jRCaMP1b.WPRE.SV40 Biosensor GENIE, Douglas Kim AV-1-PV3851...

...Screening Use pooled CRISPR libraries to screen for genes involved in specific biological processes. For more... Plasmid Pooled Libraries . ID Name Description Genes/Insert Selection PI Human knockout pooled libraries...library in backbone lentiGuide-Puro targeting 19,114 genes and containing 76,441 unique sgRNAs along with 1000...SpCas9. 76,441 unique sgRNAs targeting 19,114 human genes along with 1000 non-targeting controls Puromycin...library in backbone lentiCRISPRv2 targeting 19,114 genes and containing 76,441 unique sgRNAs along with 1000...and 76,441 unique sgRNAs targeting 19,114 human genes along with 1000 non-targeting controls Puromycin... in backbone XPR_502 (P65 HSF) targeting 18,885 genes and containing 56,762 unique sgRNAs. This backbone...

...Gene Switch: These constructs contain two genes of interest, Genes A and B. When Cre is absent, only Gene...Recombinase Plasmids Recombinase-Dependent Empty Backbones Reporter Plasmids Pooled Libraries/Kits Site-specific...manipulate DNA and control the expression of specific genes in cells or organisms. Each recombinase enzyme recognizes...have adapted these systems to activate/inactivate genes when a recombinase enzyme is present. Read on to... other recombinases , as well as useful empty backbones or reporters . Experimental Considerations Recombinase...orientation, and type of target sites that flox your genes of interest makes FLEx switch a powerful experimental...incorporate appropriate controls such as Cre-only lines to account for cellular toxicity in your system....

...Collection Synthetic Biology Plasmids Collection Empty Backbones Collection Molecular Biology Guide CRISPR Guide...periplasmic space between the inner and outer membranes. Targeting proteins to the periplasm reduces bacterial...independently control the expression of up to twelve genes using small-molecule inducers in the same E. coli... Plasmids containing easily measurable reporter genes (e.g., LacZ, luciferase, or fluorescent proteins...Bacterial one-hybrid (B1H) systems also use reporter genes to identify interactions between proteins and specific...Promoter activity Fluorescence (GFPmut3a) and luminescence (lux operon) Gram-negative bacteria Attila Karsi... Karsi 14080 pAKlux2 Promoter activity Luminescence (lux operon) Gram-negative bacteria Attila Karsi 14076...

...luciferase plasmids for gene expression assays and bioluminescent reporters.... Plasmids Luciferase Plasmid Collection Empty Backbones Expression Constructs Reporter Constructs Other...Resources Luciferase is the enzyme responsible for bioluminescence found in a variety of organisms, ranging from...effects on gene expression can be measured by the luminescence output. The most popular types of luciferase...must be delivered intracellularly to measure luminescence. Gaussia luciferase is secreted and more stable...experiment, read our Luciferase blog post . Empty Backbones Browse or search for popular empty backbone plasmids...expression of firefly luciferase with a V5 tag Kevin Janes 99283 pCW57.1-Luciferase Firefly TRE Lentiviral ...

...developmental biology. With its rapid development (embryogenesis is complete in ~3 days), short generation time...research model. With more than 70% of human disease genes having an ortholog in zebrafish, this species has...This includes plasmids for genome editing, empty backbones, fluorescent proteins, reporters, biosensors, ...selection of plasmids expressing Cas9 and empty gRNA backbones. A few examples of useful zebrafish CRISPR resources...Keith Joung Lab CRISPR/Cas9-based conditional mutagenesis in zebrafish - Wenbiao Chen Lab A CRISPR/Cas9...with fluorescent proteins to visualize and track clones of cells over time. Examples of useful tools for...cassettes encoding fluorescent proteins for tagging genes at endogenous genomic loci in zebrafish. NTR 2.0...

...Caenorhabditis elegans. Mos1 Based Transgenesis miniMos Transgenesis Kit - Erik Jorgensen Lab. This kit...development, and aging. Scientists can knock down genes by feeding worms bacteria expressing dsRNA or use...editing to easily engineer specific transgenic worm lines. The transparent body of the C. elegans hermaphrodite...worm research. This includes CRISPR, RNAi, empty backbones, fluorescent proteins, and Cre expression plasmids...Description Gene/Insert PI Plasmids Expressing Worm Genes/Inserts The table below lists plasmids that contain... include plasmids expressing Cas9, empty gRNA backbones, and constructs for endogenous tagging. Some examples...Jorgensen Lab. A collection of plasmids that streamlines the production of targeting vectors for tag insertion...

...pluripotent stem cell (hiPSC) lines. The plasmids used to generate these lines have been deposited here with...from the hiPS cell lines gRNA sequences used in the creation of the hiPS cell lines The following plasmids...proteins of key cellular structures in human cell lines. Depositor...proteins of key cellular structures in human cell lines of their choice. For a full description of how to...plasmids can be used to create cell lines with endogenously-tagged cellular structures. Addgene ID Plasmid...HIST1H2BJ-mEGFP AICSDP-52 mEGFP Histone H2B type 1-J Histones 109119 CTNNB1-mEGFP AICSDP-47 mEGFP Beta-catenin...nuclear antigen (PCNA) DNA replication foci IPSC cell lines can be ordered from the Allen Institute for Cell...

...containing adenoviral or herpesviral helper genes. These genes are not present in the AAV genome, but are... and Cap genes for packaging in specific serotypes. Some also include adenoviral helper genes (E4, E2a...helper virus genes are instead provided by helper plasmids and specific packaging cell lines. Helper plasmids...containing the AAV replication (Rep) and capsid (Cap) genes necessary for the production and assembly of viral...packaging plasmids also include adenoviral helper genes normally contained in the helper plasmid. Packaging...generally encode the adenoviral E4, E2a, and VA genes. This table contains helper plasmids in Addgene'...

...Retroviral vectors are popular for making stable cell lines or clinical use. Adenoviral Vector Guide | Adenoviral... response, useful for clinical applications as vaccines. Selecting a Viral Vector Type Each viral vector...such that they can be delivered to packaging cell lines and use their cellular machinery to generate viral...viruses. The viral genome contains only the necessary genes to achieve this goal. When a virus recognizes a ...have been stripped of any unnecessary accessory genes and divided into multi-plasmid systems. These systems...safety and are replication-incompetent. Specific genes have been mutated or deleted to prevent uncontrolled...uncontrolled viral replication, and the required genes are divided in the multi-plasmid system. Many viral plasmids...

...Institute: Patient-specific lines (Link opens in a new window) RSRT iPSC lines and isogenic controls , patient-specific... Resource Center MECP2 Animal Models Human Cell Lines CRISPR Tools Plasmids Resources The (Link opens ...global transcriptional regulator of thousands of genes and studies have suggested roles in transcriptional...almost 46% of all Rett syndrome cases. Research Milestones 1992 - MECP2 is discovered by Adrian Bird's lab...new window) PMID: 21074045 2017 - the Bird lab determines a minimally functional MECP2 that rescues neurological...opens in a new window) Envigo. The following cell lines are available directly from the labs in which they..., please email [email protected] The following cell lines are available directly from the labs in which they...

...repeat cycles of expansion. Chemokines Chemokines, or chemotactic cytokines, are small proteins that can...factor FLJ40431 Return to top Chemokines Chemokines, or chemotactic cytokines, are small proteins that can...sites of inflammation throughout the body. Cytokines Cytokines are molecules that enable cells of the immune...motif) receptor 1 CCXCR1, GPR5 Return to top Cytokines Cytokines are molecules that enable cells of the immune...Addgene’s immunology collection contains plasmids for genes involved in innate and adaptive immune signaling...response. Browse Plasmids for Immunology Research Genes have been grouped into tables based on the Gene ...plasmids for each gene. Please note that not all genes have plasmids available for request. Antigen Presentation...

...Over time, a loss of motor neurons leads to limb weakness, difficulty speaking, swallowing, and breathing...mechanisms are still poorly understood, but several genes and mapped loci seem to be associated with the disease...known genetic mutations in one of the following genes: SOD1, C9ORF72, TARDBP, FUS and OPTN. Huntington's... a mutation in one of three different inherited genes: Presenilin 1, Presenilin 2, and APP. The majority... are a risk factor for late-onset AD. Many more genes have been associated with Alzheimer's disease, through...now investigating the role that these additional genes may play in Alzheimer's disease. Research Tools ...type or mutant ORFs, gene fragments, and gRNAs for genes linked to neurodegenerative disorders. Find plasmids...

...performs a chemiluminescent reaction in the presence of several different substrates. The luminescence produced...Recombinant antibodies are created by cloning antibody genes from specific hybridoma cells into plasmids. Plasmids...region is a stable part of the antibody, which determines the isotype of the antibody. B cells, the immune...tears, saliva, mucus) where it protects mucosal membranes Dimer with a valency of 4 IgA1; IgA2 IgD Coexpressed... epitope, or polyclonal, meaning there are many clones that are specific to multiple epitopes on one antigen...Sodium azide, an antimicrobial agent, can block some amines needed for conjugation reactions and should be ... tubulin (sometimes referred to as housekeeping genes). This allows for relative comparison of expression...

...top Plasmids Encoding Mammalian Genes or Inserts Several mammalian genes have been identified as having...tables below to find popular plasmids expressing genes encoding SARS-CoV-2 proteins, (spike protein, S;... and SARS-CoV-2. Find plasmids related to these genes below. ID Plasmid Description Industry PI Additional...beating of airway cilia (Kogiso et al., 2020) Cytokines - SARS-CoV-2 infection induces a cytokine storm...storm response in the body. For a full list of cytokines and related plasmids see our Immunology Resource...Spike Receptor Binding Domain Site-Saturation Mutagenesis Libraries Screening Jesse Bloom Tyler Starr Libraries...

...identify positively or negatively selected sgRNAs, genes, or pathways. For use with genome-scale CRISPR-Cas9...CRISPR screens targeting a long or short list of genes provided by the user. Developed by the Fredrik Wermeling...tool that identifies putative target sites for S. pyogenes Cas9, S. aureus Cas9, S. thermophilus Cas9, N....mouse, rat, chicken, frog, zebrafish, fly, worm, Japanese rice fish, maize, Arabidopsis, and rice. Proof-of-concept...checks for off-target binding and can work for S. pyogenes , S. thermophilus , or N. meningitidis Cas9 PAMs...Protocol for Genomic Deletions in Mammalian Cell Lines pSpCas9(BB) (pX330) PDF, 330 KB Parrott NEBuilder...sequencing Cas9: pMLM3613 dsDNA donor plasmid backbones pSL1180-HR-PUbECFP & pSL1180polyUBdsRED PDF, 597...

...properties. Where available, we have listed cloning backbones and control plasmids. Donor Acceptor λ Dex QY ...Amber (mVenus Y67C) FRET standards and cloning backbones Koushik et al. (2006) Cerulean, Venus, and VenusY67C...FRET Positive and negative controls and cloning backbones for mTurquoise2-mVenus(L68V) Goedhart et al. (...93% Positive and negative controls and cloning backbones for various FRET pairs with mTurquoise2 Mastop...mTurquoise2 Positive and negative controls and cloning backbones for red, yellow, and turquoise FRET pairs Gadella...paired with a fluorescent protein acceptor for Bioluminescence Resonance Energy Transfer (BRET) . Browse Addgene's...

...tet-inducible vectors. Tetracycline-inducible Backbones Find backbones or constructs to use for cloning to control...endogenous systems that respond to natural stimuli like hormones or metal ions, scientists have adapted an antibiotic...interest from two separate vectors. Stable cell lines or transgenic animals can be made to achieve greater...Browse the tables below to find popular empty backbones, expression vectors for transactivators, and vectors...30 variant)-based backbone rtTA-Advanced TRE3G Johannes Zuber 60495 pSBtet-GP Sleeping Beauty transposon...Bujard, H. (1995). Transcriptional activation by tetracyclines in mammalian cells . Science, 268 (5218), 1766...

...highlights plasmids that can be used as empty backbones to express your gene of interest in plants. You...compatible with GoldenBraid. Golden Gate (MoClo) Johannes Stuttmann 1000000233 Joint Modular Cloning (JMC...gene in plants. CRISPR Sylvestre Marillonnet , Johannes Stuttmann 1000000084 pDGE Dicot Genome Editing...genome editing in dicotyledonous plants. CRISPR Johannes Stuttmann 1000000236 MoClo Plant Parts III: Transformation... reporter plasmids that feature fluorescent, luminescent, and color-producing proteins for detection of...mycorrhiza-inducible promoters to label organelles, membranes and cytoskeletal elements in M. truncatula . Return...

...fluorescent iRFP variants have been shown to have high brightness and allow for single or multicolor imaging. Photoactivatable...Far-Red Protein Excitation (nm) Emission (nm) Brightness Plasmids mCardinal 604 659 18 pcDNA3-mCardinal...Near-Infrared Protein Excitation (nm) Emission (nm) Brightness Plasmids emiRFP670 642 670 12 pemiRFP670-N1 miRFP670...Excitation (nm) Emission (nm) Activation (nm) Brightness Plasmids PAiRFP1 690 717 660 or 750 3.2 pPAiRFP1...Additional Resources Fluorescent Proteins: Empty Backbones Fluorescent Proteins: Subcellular Localization...

...membrane TdTomato IX Membranes Palmitoylation tdTomato Connie Cepko 22479 FUmGW Membranes Palmitoylation EGFP...pCAG-mGFP Membranes Palmitoylation sequence from GAP43 EGFP Connie Cepko 61099 Lck-GFP Membranes Palmitoylation...for cloning in our Fluorescent Proteins Empty Backbones Collection ). Then, by examining the overlap between...EGFP Steven Green 36209 pPalmitoyl-mTurquoise2 Membranes Palmitoylation sequence from p63 mTurquoise2 Dorus...EGFP-CAAX Endoplasmic reticulum, Golgi, and Plasma membranes CAAX EGFP Lei Lu 14873 GolgiCFP Golgi apparatus...

...will promptly notify you by email within a few business days of your request to let you know that we don...solution. You must reply to that email within 10 business days in order to proceed with any adjustments ...analysis, you will be notified by email. If Addgene determines that our current queue is too high and we do ... will notify you by email before, or within 10 business days of ordering. We will give you the option ...solution. You must reply to this email within 10 business days in order to proceed with any adjustments ...

...recombinase-expressing mouse lines 2–9 and combinations of transgenic recombinase-expressing animal lines and retro-grade...these ORF fragments (which have become exons) determines the logical expression requirements of Cre and...Flp-recombinase-expressing transgenic mouse and rat lines. The Fenno Lab maintains a public folder (Link opens...genetically-encoded calcium indicators, and rabies targeting genes. Recombinases Addgene ID Plasmid Logic Sites and...opens in a new window) Mandelbaum G, Taranda J, Haynes TM, Hochbaum DR, Huang KW, Hyun M, Umadevi Venkataraju...

...Collection for plasmids that can be used to create cell lines with endogenously-tagged gene variants or browse...human cells to pluripotency by activating enogenous genes Human pluripotent reprogramming with CRISPR activators...lentiviral plasmids Induced Pluripotent Stem Cell Lines Derived from Human Somatic Cells. Science. 2007 ...stem cells. Stem Cells. 2009 May . 27(5):1042-9. Townes Lentivirus Human Single polycistronic lentiviral...from four separate plasmids Defining molecular cornerstones during fibroblast to iPS cell reprogramming ...iPSCs Bipolar Neurons Lentiviral Human Rapid neurogenesis through transcriptional activation in human ...

...more effective than dCas9 alone for mammalian cell lines Interfere Activate (CRISPRa) Increase gene expression...can be transient, or you can generate stable cell lines if your plasmid contains a selection marker. Plasmid..., note that this method is most useful for cell lines that you can transfect at high efficiency, such ... are a popular option for producing stable cell lines, since the genetic cargo (in this case, CRISPR components...is a common choice for generating modified mouse lines. Protein delivery Lastly, you can deliver Cas-gRNA...Elongation roadblocks mediated by dCas9 across human genes modulate transcription and nascent RNA processing...

... induced pluripotent stem cell (hiPSC) lines. These cell lines were created using CRISPR and the donor...tagging transcription factors with FLAG in human cell lines. To repeat the tagging in your system, use the ...terminal affinity tag (3xFLAG-2xSTREP) on endogenous genes for the isolation of native protein complexes. This...be scaled up for systematic editing of multiple genes. Plasmids can be found associated with the following...

...collection of genes and markers tagged with fluorescent proteins as well as hundreds of empty backbones. FRET ...Fluorescent Protein Plasmids by Category Empty Backbones Study your protein of interest by fusing it to...signalling pathways. Luciferase Plasmids Use bioluminescent reporters for studying gene expression, signaling...

...modifications to the the pX458 plasmid to expand its usefulness for the lab's gene regulation and developmental...The plasmids listed here are provided as empty backbones that utilize Golden Gate cloning to insert the...E., Schneider, S. R., Abraham, J., Hensley, A., Jones, K. L., & Brzezinski, J. A. (2021). Initiation of...fate specification, yet differs from deleting both genes . Development. doi: 10.1242/dev.190272. PMID: 32541005...