We narrowed to 704 results for: des.1
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TypeCollection...LuxSit-i and neoLux1.2 : Artificial luciferases desined de novo using deep-learning approaches. Split Luciferase... Eric Campeau 21474 pLenti CMV V5-LUC Blast (w567-1) Firefly CMV Lentiviral expression of firefly luciferase...Structurally destabilized firefly luciferases : A set of three increasingly destabilized mutants which...mammalian systems. ID Plasmid Luciferase Type(s) Description PI 64784 pGL3-Basic-IRES Firefly Insertion of...plasmids. ID Plasmid Luciferase Type Promoter Description PI 60226 AAV:ITR-U6-sgRNA(backbone)-pEFS-Rluc...your regulatory element of interest. ID Plasmid Description Gene/Insert PI Return to Top Other Highlighted...
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Negative Can Be Positive: Open AAV Data with Addgene
TypeBlog Post...across scientific disciplines. Scientific Data, 8(1), 1–11. https://doi.org/10.1038/s41597-021-00981-0 Wright...excellent at infecting our target neurons (Figure 1). However, this one set of experiments didn’t provide...yet none of that data was published. Fig. 1: Testing of three different AAV serotypes (AAV1, 8...392–400. https://doi.org/10.1038/s41434-018-0028-5 de Backer, M. W. A., Brans, M. A. D., Luijendijk, M....purification and formulation. Molecular Therapy, 12(1), 171–178. https://doi.org/10.1016/j.ymthe.2005.02.021...Stimulation. Frontiers in Human Neuroscience, 14(December), 1–12. https://doi.org/10.3389/fnhum.2020.578687 Additional...of negative data. Negative AAV data is highly desirable In a survey we conducted to improve our Addgene... -
Addgene-Seeding Labs Plasmid Grant: Accelerating Science Globally - 5 Scientists, 4 Countries & 3 Continents!
TypeBlog Post... compounds with various antimicrobial activities (1). She has developed a fast endophyte screening system...Rodrigo Arturo Rivera Solís, Universidad Autónoma de Yucatán, Mexico Dr. Rodrigo Arturo Rivera Solí...us coordinate and promote the awards. References 1. Martinez-Klimova, Elena et al. “Endophytes as sources...antibiotic” Biochemical Pharmacology 2017 Jun 15;134:1-17. PubMed PMID: 27984002. 2. Bilan, Dmitry S. et ...renewable biotechnology products and biosensors for herbicides as well as developing yeast as source for dietary...technology to engineer plants that are able to destroy DNA viruses, similar to the anti-phage CRISPR effect...cloning kit) she intends to make yeast capable of desaturating fatty acids. Identification of Insecticidal ... -
Targeting HIV-1 with CRISPR: Shock and Kill or Cut it Out?
TypeBlog Post...plasma HIV-1, improving life expectancy and quality for HIV-1 patients. However, ART is not an HIV-1 cure. ...activated HIV-1, measured via HIV-1 promoter-driven GFP expression. In cell lines that can produce HIV-1 toxic...with HIV-1. For two HIV-1 strains, Cas9/gRNA lentiviral expression significantly reduced the HIV-1 copy number...HIV-1 transcription and destroy infected cells, the other using wild-type Cas9 to remove the HIV-1 genome...currently infected with the lentivirus HIV-1. Today, HIV-1 can be controlled with antiviral therapies ...take on HIV-1 in vitro, and what obstacles must be overcome for clinical success. ART and HIV-1 reservoirs...Cutting HIV-1 out of the genome with CRISPR/Cas9 Before the introduction of CRISPR, HIV-1 genome editing... -
CRISPR References and Information
TypeCollection..., 2.4 MB Zhang GeCKO pooled library amplification 1 vector system: lentiCRISPR v2 2 vector system: lentiCas9...CRISPR/Cas design (Link opens in a new window) Program for designing optimal gRNAs. Provides feedback on...window) This tool helps design (10 different prediction scores), clone (primer design), and evaluate gRNAs...Addgene Church gRNA design and cloning gRNA cloning vector PDF, 118 KB Church gRNA design and cloning for ...window) CRISPR/Cas9 reagent generation; gRNA design; HDR design; Deep sequencing Cas9: pMLM3613 dsDNA donor...CRISPR resources and references, including gRNA design tools, CRISPR FAQs, and community forums. CRISPR...protoSpaceJAM is an all-around platform for CRISPR knock-in design at scale. It uses an open-source algorithm to automate... -
COVID-19 Resources
TypeCollection...mediating the entry of SARS-CoV and SARS-CoV-2 viruses. ( 1 ) TMPRSS2 - a serine protease that primes the SARS-CoV...SARS-CoV-2 Viral Pseudotyping Collection - This page includes a table of all spike expression plasmids in our... industry is continually growing. ID Plasmid Description Industry PI Return to top Other Coronavirus Plasmids... SARS-CoV, MERS, BAT-CoV Plasmids ID Plasmid Description Industry PI Due to export control regulations...plasmids related to these genes below. ID Plasmid Description Industry PI For more information on these genes...CRISPR-Cas13: Open-access SHERLOCK research protocols and design resources (Link opens in a new window) Find Zhang...COVID-19 research in the table below: ID Plasmid Description Industry PI Do you have a suggestion for plasmids... -
Crowdfight, a Platform to Boost Scientific Collaboration During COVID-19 and Beyond
TypeBlog Post...postdoctoral researcher at Instituto de Investigación Biomédica de Salamanca (IBSAL) and one of the coordinators...prompted us to create Crowdfight. Figure 1: Crowdfight is a community of scientists who believe...collaborator for a specific task, Crowdfight will: 1) Find an expert with the right skill; 2) Confirm the... illustrates the power of collaboration, which despite the competitiveness of cutting-edge research, is...physics. Even non-scientists, such as lawyers, designers and other professionals, joined the platform; ... -
Designer PUF Proteins for Any RNA Target
TypeBlog Post...regulation of endogenous RNA. J Biol Eng. 2014 Mar 1;8(1):7. doi: 10.1186/1754-1611-8-7. PubMed. Related Resources...targeting specific sequences of RNA is highly desirable for extending the complexity of genetic circuits...Zhao and colleagues, we now have just the tool…designer PUF proteins! A newly available PUF Assembly Kit...In a single step, researchers can now assemble designer PUF domains for RNA specificity engineering. “...proteins The tool she and her colleagues led by Zhao described and demonstrated in a report published in The ...translational repressor. In principle, however, designer PUF proteins can be fused to other components ...standard BsaI in place of the high fidelity version (despite what the original paper says). In their hands, ... -
Antibody Neutralization Response Against Pseudoviruses Expressing SARS-CoV-2 Spike Protein Variants
TypeBlog Post...for the wild-type. However, variants such as the P.1 (Gamma variant) and the B.1.351 (Beta variant) were... tested at the time of this study. Figure 1: Neutralization levels against spike pseudovirus decreases...lowest levels of cross-neutralization occurs in the P.1 and B.1.351 variants tested. Image from Garcia-Beltran..., Gosnell BI, Hanekom W, Moosa M-YS, Lessells RJ, de Oliveira T, Sigal A (2021) Escape of SARS-CoV-2 501Y.V2... -
An Introduction to Adenovirus
TypeBlog Post...unusual cases was either 1) a physiological effect of SARS-CoV-2 toward destabilizing one’s natural immune...an AdV transfer plasmid with the E1 and E3 (Early 1 and Early 3) genes deleted. These genes’ functions...certain contexts (Bauzon et al., 2003). Figure 1: The adenovirus life cycle. Image reused from Kremer... review of the evidence. Emerg Microbes Infect, 8(1), 1679–1687. https://doi.org/10.1080/22221751.2019.1690953...adenoviral vector infected cells does not require de novo viral gene expression: Implications for gene...574–580. https://doi.org/10.1038/s41586-023-05949-1. PMID: 36996871. Tatsis, N., Fitzgerald, J. C., Reyes-Sandoval...viruses infecting humans (from seven species, designated as letters A–G) exist. The life cycle of AdVs... -
Antibody Guide
TypeCollection...and placed in a buffer. Antibody Structure Figure 1: Structure of an antibody A standard antibody is made... together to form a “Y” shape, as shown in Figure 1. The two arms of the Y structure are responsible for... is described in more detail in the Multiplex Assays section below. The following table describes the ... Science Guides Antibody Guide Antibody Guide View Addgene's Antibody...of B cells isolated from animals immunized as described above are used to create hybridomas, which produce...each antibody has two binding sites. Isotype Description Configuration and Valency Subclasses IgA Prevalent... used to quantify relative amounts of protein. Includes: Western blot ELISA Capture - used to isolate ... -
Split Fluorescent Proteins for Studying Protein-Protein Interactions
TypeBlog Post... help! General Design Figure 1: Two proteins are fused to the FP(11) and FP(1-10) fragments. When... Color Base FP Description Article PI Blue EBFP2 EBFP2(1-10) and Capri(1-10) for use with GFP...and 11 to yield large (1-10) and small (11) fragments. In several cases an FP(1-10) fragment may be compatible...Commun. 2017 Bo Huang mNeonGreen3 mNG3K(1-10) and mNG3A(1-10) for use with mNG2(11) Improved yellow-...sfCherry2 sfCherry2(1-10) and sfCherry2(11), also photo-activatable variant PAsfCherry2(1-10) Improved split...brings the pieces of the FP back together (Figure 1). This method is generally known as Bimolecular Fluorescence...even from a different base FP (for example, EBFP2(1-10) can be used with GFP(11)). Some of the articles... -
pCXLE toolkit: Efficient episomal plasmid-based method to reprogram peripheral blood cells to iPSCs
TypeBlog Post...replication, OriP and the EBNA-1 sequence that encodes for the DNA binding protein EBNA-1 [7]. These two sequences...are based on the Epstein-Barr Nuclear Antigen-1 (oriP-EBNA-1) system where each vector contains a viral ...extra plasmid, pCXWB-EBNA-1 is used in the reprogramming cocktail. pCXWB-EBNA-1 lacks oriP and is unable...expressing additional EBNA-1 transiently. The transiently expressed EBNA-1 increases protein expression...pCXLE vectors listed below in Table 1 are available at Addgene. Table 1: List of pCXLE episomal plasmids,...obtained and the subsequent reprogramming success [1]. Fibroblasts need to be cultured and expanded in ...reprogramming factor cocktail, and viral protein EBNA-1. Methods for reprogramming hiPSCs from PBMCs While... -
Tips for a 1st time AAV user (by a Rookie AAV user)
TypeBlog Post...different plasmids to produce AAV: 1) the RepCap plasmid which provides the AAV replication (rep) and capsid...plasmid. This whole process takes ~3 hours to complete: 1 hours hands-on time and 2 hours for the qPCR run and... Special thanks for Dr. Dan Stone and Harshana de Silva Feelixge at Fred Hutchinson Cancer Center for...about AAV production and purification! References 1. Aurnhammer, C., Haase, M., Muether, N., Hausl, M.... repeat sequences. Human gene therapy methods, 23 1, 18-28. PubMed PMID: 22428977. 2. Choi, V.W., Asokan...replication; and 3) the transfer plasmid which encodes a transgene of interest that’s packaged into the... -
Starter Guide to induced Pluripotent Stem Cells (iPSCs) Part 2: Reprogramming and Transdifferentiation
TypeBlog Post...type of choice in vitro is known as reprogramming [1]. The process can be divided into two stages: Dedifferentiation...//www.linkedin.com/in/kmukherjeephd/. References 1. Hochedlinger, K. and R. Jaenisch, Nuclear reprogramming...from human peripheral blood. Cell Stem Cell, 2010. 7(1): p. 15-9. PubMed PMID: 20621044. PubMed Central PMCID...induced pluripotent stem cells. Hepatology, 2010. 51(1): p. 297-305. PubMed PMID: 19998274. PubMed Central... pluripotent stem cells. Nat Biotechnol, 2014. 32(1): p. 84-91. PubMed PMID: 24291815. PubMed Central ...keratinocyte lineage. Methods Mol Biol, 2014. 1195: p. 1-12. PubMed PMID: 24510784. PubMed Central PMCID: PMC4096605...induced pluripotent stem cells. PLoS One, 2010. 5(1): p. e8763. PubMed PMID: 20098701. 31. Tang, Z.H.,... -
Plasmids 101: Common Lab E. coli Strains
TypeBlog Post...Table 1 below outlines a few of the more common genetic changes found in E. coli strains. Table 1: Common...amber (UAG) stop codon by tyrosine insertion λ-thi-1 or thi1 Mutation in thiamine metabolism Requires exogenous... all based on E. coli K-12 and are considered BSL-1. Table 2: Lab strains of E. coli Strain Natural...plasmids, blue/white screening. F- endA1 glnV44 thi-1 recA1 relA1 gyrA96 deoR nupG Φ80dlacZΔM15 Δ(lacZYA-argF...) recA13 leuB6 ara-14 proA2 lacY1 galK2 xyl-5 mtl-1 rpsL20(SmR) glnV44 λ- JM109 General cloning and...strain for cloning repetitive DNA. endA1 glnV44 thi-1 relA1 gyrA96 recA1 mcrB+ Δ(lac-proAB) e14- [F' traD36...of an E. coli K-12 strain. F- λ- ilvG- rfb-50 rph-1 NEB Stable For cloning into and storage of lentiviral... -
Great Results Start with Great Standard Curves
TypeBlog Post... Figure 1: A 1 mg/mL control is diluted in a series ranging from 1:2 to 1:32. Each individual...example, Figure 1 depicts a 2-fold dilution series. Each step of the series dilutes by 1:2 for a series...series that ranges from 1:2–1:32. The specific dilution series used will depend on the expected concentration... points fit the trendline. R2 ranges from 0 to 1 with 1 being a perfect fit and 0 indicating that there...individual step of the series is consistent at 1:2. When preparing a dilution series, use a new pipette tip for... the data points generated by replicate values provides useful information about the accuracy of the curve...If the unknown sample’s measurement is too low, redesign the standard curve dilution series with lower ... -
CRISPR Between the Genes: How to Experiment with Enhancers and Epigenomics
TypeBlog Post...three-step approach for designing an experiment to identify the critical NANOG binding sites: 1. Find Enhancers...in combination with the techniques from Table 1, provides us with powerful GPS-like techniques to explore... chromosome, start, and stop. For instance (chr3, 1, 10) indicates a stretch of DNA at the very beginning...genetic elements like protein, RNA, and DNA (Table 1). As with GPS in the real world, coordinates alone...genomic coordinates into meaningful experiments. Table 1: An emerging family of "GPS for the genome" techniques...enhancers with the help of genome math. Figure 1: Genome math illustrated. Intersect (left) and set...histones with “GPS for proteins,” ChIP-seq from Table 1. Because of DNA’s 3D geometry and the chemical properties... -
Celebrating the 2024 Nobel Prize in Chemistry
TypeBlog Post...protein design goals, both from the Baker Lab and others. The introduction of de novo designed proteins...protein structures, called Top7. Figure 1: Top7 receives the Nobel Prize. Created with BioRender.com...concept that this kind of protein design is possible. Although Top7 was designed simply to be stable and novel...Hassabis and John Jumper! The Baker Lab focuses on designing entirely new proteins, improving our understanding...team was the first to develop and publish a human-designed protein that was not based on any existing protein...least that we’ve seen so far!). They started by designing a novel topology, then iteratively generated 3D...got closer and closer to achieving it. Once they designed the protein computationally, the researchers expressed... -
Lambda Red: A Homologous Recombination-based Technique for Genetic Engineering
TypeBlog Post...recombineering system has three components (Figure 1): 1) Exo, 2) Beta, and 3) Gam. All three are required...frequency between 0.1% to 1%, and can be increased to as high as 25-50% by designing oligos that avoid activating...activating MMR: 1) use a strain of bacteria that has key MMR proteins knocked out or 2) specially design ssDNA ... These changes can be 5’ or 3’ of the desired change. Table 1: Overview of ds- and ss-DNA Uses in Recombineering...Considerations dsDNA Insertion or deletion of >20 nucleotides 1 out of 104 to 105 Use a high fidelity Taq polymerase... ssDNA Point mutations or changing a few nucleotides 0.1-1% or 25-50% when activation of MMR is avoided...from the 5’ end and generate 2 possible products: 1) a partially dsDNA duplex with single-stranded 3’ ...