...types of luciferase found in Addgene’s collection are Firefly luciferase (Fluc), Renilla luciferase (Rluc...and NanoLuc® Luciferase (Nluc). Luc2 is the second-generation version of Firefly luciferase that has been... Plasmid Collections Luciferase Plasmids Luciferase Plasmid Collection You may also like...Fluorescent Proteins Blog: Luciferase 101 Blog: Technologies Enabled by NanoLuc® Luciferase Blog: Luminescent ...mammalian systems. Other luciferases in our collection include Gaussia luciferase (Gluc) and the red and...green Click-beetle luciferases (CBRluc and CBGluc, respectively). Like Firefly luciferase, Click-beetle red...type of luciferase has advantages and disadvantages depending on the application. Firefly luciferase is considerably...

...Fluorescent Proteins Other Resources Neurodegenerative Disorders Neurodegenerative diseases are a group of disorders... the progressive degeneration and ultimate death of neurons. Common neurodegenerative diseases include...most neurodegenerative diseases are only partly understood, and there are no effective therapies to prevent...available at Addgene. Amyotrophic Lateral Sclerosis Amyotrophic lateral sclerosis (ALS), sometimes referred...Addgene that may be useful for neurodegeneration research. Neurodegeneration... Neurodegeneration Research Collection Addgene ...plasmid collections that may help with your neurodegeneration research. Disease Info Plasmid Collection ...

... Neurodegeneration Neurodegeneration Plasmids Neurodegeneration Plasmid Collection ... 54155 mEmerald-MAPTau-N-10 MAPT mEmerald CMV Parkinson's, FTD Michael Davidson 54226 mEmerald-Peripherin-C...mEmerald-Peripherin-C-14 PRPH mEmerald CMV ALS Michael Davidson 54227 mEmerald-Peripherin-N-18 PRPH mEmerald CMV ALS Michael... on genes and associated neurodegenerative diseases was curated from several sources (Chan Zuckerberg ...Parkinson's John Chodera 79712 E2AK2_HUMAN_D0 EIF2AK2 His, TEV T7 VWM disease John Chodera 82285 pDONR223...Lippincott-Schwartz 164210 pLEX-EF1a ANXA11-mEmerald ANXA11 mEmerald EF1a ALS Jennifer Lippincott-Schwartz ...Lippincott-Schwartz 164211 pLEX-EF1a ANXA11-mEmerald D40G ANXA11 mEmerald EF1a ALS Jennifer Lippincott-Schwartz 164212 pLEX-EF1a...

...Find plasmids generated by the iPSC Neurodegenerative Disease Initiative (iNDI) here....Stem Cell Research Neurodegeneration Research Collection The iPSC Neurodegenerative Disease Initiative...Collections iNDI iPSC Neurodegenerative Disease Initiative (iNDI) Collection You May Also Like... CRISPR ...Initiative (iNDI) project aims to generate and characterize single nucleotide variant lines in an isogenic cellular...Addgene. Figure 1: Schematic of iNDI iPSC line generation, phenotyping, and distribution efforts. Plasmids...NIH. Reference Ramos et al. 2021. Tackling neurodegenerative diseases with genomic engineering: A new stem...

...Cas-OFFinder and Microhomology predictor. CRISPR-ERA: CRISPR -mediated e diting, r epression, and a ctivation...CRISPR Discussion Group (moderated by Feng Zhang lab): Forum/FAQ Drosophila: General Advice and Forum/FAQ ...entries describing enzyme activity, experimental considerations, and more. CasPEDIA is a community resource... KB Vosshall and Matthews CRISPR/Cas9 reagent generation; gRNA design; HDR design; Deep sequencing Cas9...

... DNA Polymerase I 165533 pOpen-TthpolA Tth DNA polymerase 165534 pOpen-T7gene5 T7 DNA Polymerase (unmodified... Ligases DNA Polymerases Restriction Endonucleases Reverse Transcriptases RNA Polymerases Other Enzymes... of molecular biology, including DNA polymerases, RNA polymerases, DNA ligases, reverse transcriptases... DNA Polymerases Addgene ID Plasmid Gene/insert 165500 pOpen-9N7polA 9 degrees N-7 DNA polymerase 165501.... Heat-Stable DNA Polymerase 165502 pOpen-9N7polA (CT) 9 degrees N-7 DNA Polymerase chain terminating ...pOpen-K12polLF DNA Polymerase I, Large (Klenow) Fragment 165522 pOpen-K12polLF (exo-) DNA Polymerase I, Large (...

...channel Mouse Chimera: Mouse/Rat rat IgG2a 222182 RGS14 [N133/21R-rat IgG2a] RGS14 Rat Chimera: Mouse/Rat...receptor Rat Chimera: Mouse/Rat rat IgG2a 222188 GFP [N86/8R-rat IgG2a] GFP Aequorea victoria Chimera: Mouse... to the research community. Protocols for the generation R-mAbs plasmids, production of R-mAbs in transfected...researchers will be able to easily and inexpensively generate R-mAbs by transiently transfecting the plasmids...channel [K14/16R-rat IgG2a] Kv1.2 K+ channel Human Chimera: Mouse/Rat rat IgG2a 222178 CASPR/Neurexin IV [... [K65/35R-rat IgG2a] CASPR/Neurexin IV Rat Chimera: Mouse/Rat rat IgG2a 222179 GluA2/GluR2 glutamate receptor...-rat IgG2a] GluA2/GluR2 glutamate receptor Rat Chimera: Mouse/Rat rat IgG2a 222180 Slo1/BKAlpha maxi-K...

...collection of plasmids for the generation and study of stem cells, from iPSC generation to directed differentiation...source for therapeutic use. Since 2006, scientists have developed many approaches to generate iPSCs. The... post Delivery Methods for Generating iPSCs provides an overview of several different reprogramming methods...Non-viral Method to Generate Integration-Free Human iPS Cells from Cord Blood and Peripheral Blood Cells. Stem... Stem Cell Research Plasmids Stem cells are proliferative, unspecialized cells with the capacity to differentiate...pluripotent or multipotent, based on their ability to generate different cell lineages. Pluripotent stem cells... for drug development, disease modeling, and regenerative medicine. iPSCs, much like their ES cell cousins...

...using dual nickases to generate a knockout or gene edit; or using Cas9 to generate large genomic deletions...proteins. Large deletions generated by Cas3 have fewer off-target alterations, such as small mutations ...nucleases (TALENs) required scientists to design and generate a new nuclease pair for every genomic target. ..., and precisely edit DNA and RNA. The ease of generating gRNAs makes CRISPR one of the most scalable genome...and some of the ways the CRISPR components are leveraged to achieve specific goals, such as targeting multiple...depends on which Cas protein you use, and there are several to choose from. Many variations of Cas9 exist, ...will form a ribonucleoprotein complex through interactions between the gRNA scaffold and surface-exposed...

...signaling molecules are horseradish peroxidase (HRP) and fluorophores. Horseradish peroxidase (HRP) HRP ...with several suitable antibodies; if they all meet your criteria, you may either purchase several to compare...epitopes on one antigen. Monoclonal antibodies are generated from cultured B cells, where each culture should...variability. Other Affinity Reagents There are several alternatives to antibodies, ranging from antibody... aliquots of at least 10 µL. Antibodies cannot tolerate freeze-thaw cycles. Here are a few tips for storing...a chemiluminescent reaction in the presence of several different substrates. The luminescence produced...Applications Antibody-based applications can be generally classed into four categories: Quantification -...

...The Rinehart Lab has developed several tools useful for the incorporation phosphoserine into your gene...phosphoprotein yield, rEcoli growth characteristics, and an overall improved ease of use when expressing recombinant... in Mohler et al., Mol Syst Biol 2023 and find general tips on phosphoprotein expression in rEcoli XpS... Biotech. 2018). This phosphosite library was generated in a single mixed pool by expressing and purifying...assessments. The mode #1 phosphosite library can be generated with either phosphoserine using SepOTSλ ( 68292...screening phosphorylation-dependent protein-protein interactions. The phosphosite DNA library can also be introduced...of phosphorylation-dependent protein-peptide interactions (“mode #2” expression, Barber et al, Nat. Biotech...

...and generate virus (right), similar to how natural viruses can be delivered to a host and generate virus... Adenovirus vectors for gene therapy, vaccination and cancer gene therapy. Curr Gene Ther. 2013 Dec;13...can direct long-term transgene expression, but generally are not permanently integrated into the host genome...oncolytic) adenoviruses are utilized for cancer therapy because of their ability to preferentially infect...can be used to target specific cells or tissues. Overall, viruses can efficiently get genetic material into...an organism. Figure 1. Using viral plasmids to generate virus. A) Viral plasmids have been engineered ...genetic material (termed the viral genome) that, in general, encodes for more viruses. When a virus infects...

...to measure protein-protein interactions; if two proteins of interest interact in a bacterial two-hybrid...pTac GFP, Luciferase (LuxCDABE) Attila Karsi Broad host range vectors for expressing luciferase and fluorescent...to a particular sequence? Do these two proteins interact? What happens when I overexpress this enzyme at... redirecting metabolic pathways to produce a therapeutic compound, there are many different ways to alter...appropriate construct, grown at the permissive temperature (32°C) to allow transposition, and the construct... then cured by growing at the non-permissive temperature (42°C). Transposition is efficient enough that...about your protein’s function, structure, and interactions! Plasmid ID Promoter Epitope Tag(s) ID Purpose...

...used with 2nd and 3rd generation lentivirus technologies. ID Plasmid Generation Type Description PI 12253...transgene expression. There are some safety considerations to evaluate when working with lentivirus: these...potential for insertional mutagensis. The 2nd and 3rd generation lentiviral systems available at Addgene have ...information about: Lentiviral components 2nd versus 3rd generation lentiviral systems Viral production Frequently...produce an infectious viral particle (a virion) are generally divided among multiple plasmids. Some plasmids... Packaging Contains Rev; can be used with 3rd generation transfer plasmids along with additional packaging...Packaging Contains Gag and Pol; can be used with 3rd generation transfer plasmids along with a plasmid encoding...

...want to: Generate complete and permanent loss of gene expression or function ( knockout )? Generate a specific... Edit Generate a specific user-defined sequence change in a particular gene, such as generating a point...like never before. This section will provide a general framework to get you started using CRISPR in your...Manipulation Application Cas9 gRNA Additional Considerations Knockout Permanently disrupt gene function ... enrich positive cells, or selection marker to generate stable cell lines (for details, see Plasmids 101...inserted into an AAV transfer vector and used to generate AAV particles (for details, see our AAV Guide ...Cas9 and gRNA In vitro transcription reactions generate mature Cas9 mRNA and gRNA, which are then delivered...

...curated literature information (Link opens in a new window) OMIM MECP2 - MECP2 curated literature information...MECP2 reactivation, RNA editing, gene replacement therapy, RNA trans-splicing, and protein replacement to...contrast to the sustained loss of skills in neurodegenerative conditions, the regressive period in Rett ...locally or globally. The MECP2 protein contains several functional domains, including: the N - t erminal...acid and Thyroid hormone receptor (NCoR/SMRT) interacting domain (NID) a nuclear localization signal (not...the MECP2 protein in mouse and rat models have accelerated our understanding of Rett syndrome at the molecular...They are also an important tool for developing therapeutic strategies to treat Rett syndrome and restore...

...expressing firefly luciferase. pCMV-FLuc - Retroviral reporter vector expressing firefly luciferase. pHAGE-CMV-Luc2... vector expressing firefly luciferase and ZsGreen. NL4-3 mCherry Luciferase - Lentiviral dual reporter...EGFP. Lenti-luciferase-P2A-Neo - Lentiviral reporter vector expressing firefly luciferase with neomycin...poorly understood but are critical for developing therapeutic strategies to prevent viral spread. As a Biosafety...the live virus. Alternatively, it is possible to generate pseudovirus, where the structural and functional...spike protein is inefficient for pseudotyping; several modifications may enhance infectious particle production...lists expression plasmids that are suitable for generating pseudotyped virus for SARS-CoV-2 and other coronaviruses...

...the revolutionary dynamic picture of the brain generated through this effort will provide novel ways to...Hillel Adesnik 111066-AAV1 pAAV-hSyn-dLight1.1 To generate Adeno-Associated viruses for expression of dLight1.1...promoter Lin Tian 111067-AAV1 pAAV-CAG-dLight1.1 To generate Adeno-Associated viruses for expression of dLight1.1...promoter Lin Tian 111067-AAV5 pAAV-CAG-dLight1.1 To generate Adeno-Associated viruses for expression of dLight1.1...promoter Lin Tian 111068-AAV1 pAAV-hSyn-dLight1.2 To generate Adeno-Associated viruses for expression of dLight1.2...promoter Lin Tian 111068-AAV5 pAAV-hSyn-dLight1.2 To generate Adeno-Associated viruses for expression of dLight1.2...promoter Lin Tian 111069-AAV1 pAAV-CAG-dLight1.2 To generate Adeno-Associated viruses for expression of dLight1.1...

...systematic protein interaction screens or cellular assays to identify novel interactions or pathways that...method to detect interactions is AVEXIS (for Avidity-based extracellular interaction screen) in which ...protocol description: Avidity based Extracellular Interaction Screening (AVEXIS) for the scalable detection...of low affinity extracellular receptor-ligand interactions. Kerr SJ and Wright GJ. J. Vis. Exp. (61), e3881...Construction of a large extracellular protein interaction network and its resolution by spatiotemporal ...proteomics 2010 Dec; 9(12):2654-65. A cell surface interaction network of neural leucine-rich repeat receptors...for novel low-affinity extracellular protein interactions. Bushell KM, Söllner C, Schuster-Boeckler B,...

...EGFP Ken-Ichi Takemaru 54234 mEmerald-PLK1-N-16 Centrosome Plk1 mEmerald* Michael Davidson 54491 mCherry-Lifeact...Vladislav Verkhusha 79990 pKeratin-miRFP703 Intermediate Filaments (epithelial) Keratin miRFP703 Vladislav Verkhusha...mCherry Alpha Yap 54219 mEmerald-Paxillin-22 Focal Adhesions paxillin mEmerald* Michael Davidson 15233...GFP Bo Huang 54040 mEmerald-Clathrin-15 Clathrin-coated vesicles Clathrin mEmerald* Michael Davidson 79997...EEA1 GFP Silvia Corvera 42635 TagRFP-T-EEA1 Early endosomes EEA1 TagRFP-T Silvia Corvera 38770 pEF.myc....Optogenetics Fluorescent Protein Blog Posts Background General Cytoskeletal Junctions Secretory Pathway Background...structures as these well-characterized proteins. General Structures The cell contains a variety of organelles...