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  1. Plasmids 101: Using Transposons in the Lab

    Type
    Blog Post
    ...move antibiotic resistance genes between plasmids or from plasmids to the bacteria’s genome. In humans, if...which is a big advantage over viral vectors (~5 kB payload for AAV and ~8 kB payload for lentivirus). ...in viral vectors, but easily fit in a TE. Transposons are also less likely than viral vectors to induce...transposon’s LTRs, similar to when packaging viral vectors.   Let’s take a closer look at three different ... in human cells at rates similar to retroviral vectors. The hyperactive version of the SB transposase,...activity (Voigt et al., 2016). Sleeping Beauty plasmids  piggyBac Although its name suggests otherwise...efficiency in human cells similar to retroviral vectors. The hyperactive PB transposase (hyPB) is ~10-fold...

  2. Addgene Receives NIH BRAIN Initiative Grant to Create Open-Access Recombinant Antibody Resource

    Type
    Blog Post
    ...Would you buy and use a plasmid if it didn’t include any plasmid sequence data? Of course not. Unfortunately...depositing your plasmids encoding antibodies, ScFvs, and nanobodies to Addgene! The plasmids will be made...distributions of research materials, such as plasmids and viral vectors. However, it typically takes a few years...Plückthun, 2015a, b). Recombinant antibodies are plasmid-based and offer numerous advantages, including ...antibodies that you have come to expect from our plasmid and viral services. The first step will be working...working with the community to build a resource of plasmids encoding recombinant antibodies as well as other...crowdsourced Data Hub (built off of the concept of our AAV Data Hub) to share further validation data with you...

  3. Hot Plasmids: Winter 2025

    Type
    Blog Post
    ...of the new plasmids, antibodies, and viral preps in the repository through our Hot Plasmids articles.  ...et al., 2024. The authors have deposited AAV plasmids for Cre-dependent and -independent versions ...ONE vector G-protein Optical (ONE-GO) biosensors (Janicot and Maziarz et al., 2024). Each plasmid expresses...selectivity. With 10 plasmids available as a kit, plus 21 optional accessory plasmids, the collection targets...precise plant breeding. Find dicot prime editing plasmids here! Vu, T. V., Nguyen, N. T., Kim, J., Song... proteins for mammalian cell expression. These plasmids will be useful not only for future studies of ...proteins of interest too! Find CLIP, CLAP, and SNAP plasmids here! Smith, E.R., Turner, E.D., Abdelhamid, ...

  4. Treating Muscular Dystrophy with CRISPR Gene Editing

    Type
    Blog Post
    ...stem cells 2. Remove any immunogenicity of the AAV vector Once delivery has been optimized, it’s important...for future preclinical and clinical work. Find Plasmids for Disease Research on Our Special Collections...culture, but Ran et al. have shown that CRISPR and AAV can be used together for postnatal genome editing...facets of CRISPR DMD therapy. Long et al. show that AAV-CRISPR does not cause obvious germline modification... Check out Our CRISPR Guide Pages Find CRISPR Plasmids for Your Research Find Validated gRNAs for Your...with AAV9, whereas Nelson et al. and Tabebordbar et al. used the shorter SaCas9 with AAV8 and AAV9, respectively...

  5. Cpf1: A New Tool for CRISPR Genome Editing

    Type
    Blog Post
    ...deliver in low-capacity vectors, such as adeno-associated viral (AAV) vectors. In 2017, Zetsche et al....Francisella Cpf1 (FnCpf1), they employed an E. coli plasmid depletion assay to discover FnCpf1’s PAM sequence...system." Cell (2015). PubMed PMID: 26422227. Find plasmids from this publication at Addgene. 2. Zetsche,....1 (2016): 31-34. PubMed PMID: 27918548. Find plasmids from this publication at Addgene. 3. Kim, Daesik...Biotechnol. (2016). PubMed PMID: 27272384 Find plasmids from this publication at Addgene. Kleinstiver...Biotechnol. (2016). PubMed PMID: 27347757. Find plasmids from this publication at Addgene. Makarova, Kira...PMID: 26411297. Resources at Addgene Find CRISPR plasmids at Addgene Read Addgene's CRISPR Resource Guide...

  6. Hot Plasmids - February 2022

    Type
    Blog Post
    ...subset of the new plasmids and viral preps in the repository through our hot plasmids articles. These articles...recent plasmid deposits and we hope they'll make it easier for you to find and use the plasmids you need...need. If you'd ever like to write about a recent plasmid deposit please sign up here.  Here's what you'...Ladder  by: Rachel Leeson This simple, inexpensive plasmid-based ladder system is a cost-effective alternative... ladders. Image from Santilli, et al. 2021 Plasmids are used to express 10, 15, 20, 30, 40, 50, 60...50 mLs) or through two polycistronic expression vectors (3570 lanes/50 mLs) coexpressing a set of either...kD proteins or 20, 40, 60, and 80 kD proteins. Plasmids containing 150 or 250 kD proteins are also available...

  7. Multifunctional Neural Fibers: Modulation from Gut to Brain

    Type
    Blog Post
    ... BRAIN initiative plasmid collection More resources on the Addgene blog Plasmid tools for microbiome... of a live animal. They started by delivering an AAV construct expressing channelrhodopsin 2 (ChR2), a...studies Channel rhodopsins for optogenetics Viral Vectors 101: Chemogenetics ...

  8. CRISPR 101: Anti-CRISPR Proteins Switch Off CRISPR-Cas Systems

    Type
    Blog Post
    ...acids) means they can be delivered in vivo using AAV vectors. Having a way to shut off CRISPR-Cas systems ...Papayannopoulou T, Lieber A (2018) HDAd5/35++ Adenovirus Vector Expressing Anti-CRISPR Peptides Decreases CRISPR...CRISPR   Resources on Addgene.org Find CRISPR Plasmids for Your Research Interested in light-inducible...light-inducible Cas9 activity? Check out these these plasmids Catch Up on Your CRISPR Background with Our Guide Pages...

  9. Antibodies 101: Single Chain Fragment Variables (scFvs)

    Type
    Blog Post
    ...size also allows for their delivery by viral vectors like AAV, which could be useful for delivering scFvs...insect cells, they are most often expressed from a plasmid in bacteria (Ahmed et al., 2012). One challenge...domains of the selected scFvs are inserted into a vector encoding the antibody constant domain scaffold ...doi.org/10.1038/nbt.1791  Deal CE, Balazs AB (2015) Vectored antibody gene delivery for the prevention or treatment...Resources on Addgene.org: Browse Addgene’s Antibody Plasmid Collection Find antibodies for neuroscience research...

  10. Generating Mouse Models Using CRISPR/Cas9

    Type
    Blog Post
    ...or GONAD (Gurumurty et al., 2016). AAV can also be used as a vector to deliver CRISPR/Cas9 either through... Find CRISPR Plasmids for use in Mammals Check out Our CRISPR Guide Pages Find Plasmids for Mouse Targeting...the target site, and a donor oligonucleotide or plasmid as the repair material if needed (for knock in ..., the gRNA, Cas9, and donor oligonucleotide or plasmid components are brought together and microinjected...provided (in the form of an oligonucleotide or plasmid), precise changes can be made in the genome via...mice carrying the intended mutation. With a donor plasmid, the outcome is less predictable. In our best case...Mediated Genome Editing with Adeno-Associated Viral Vector. iScience 9:286–297 . https://doi.org/10.1016/j.isci...

  11. Lentivirus ddPCR Titration

    Type
    Protocol
    ...ddPCR) for lentivirus vector titration.... Protocols ddPCR Titration of Lentivirus Vectors ddPCR Titration of...of Lentivirus Vectors You may also like... Lentivirus Colony Formation Titration Assay Lentivirus Fluorescence...Fluorescence Titration Assay AAV Titration by qPCR Addgene Viral Service Lentivirus Guide Virus Blog Posts...droplet digital PCR (ddPCR) to titer lentivirus vectors. This protocol specifically uses primers and probes...green) and negative (black) droplets Sample ID Plasmid ID Dilution Copies/20 µL RRE (FAM) Copies/20 µL...

  12. Plasmids 101: The protein expression toolbox

    Type
    Blog Post
    ...some permanent knock out plasmids? Check out our Cre vector and Murine LoxP vector.   Turn it off for a little... a gene (cDNA) can be done by either plasmid or virus. A plasmid carrying your cDNA can be targeted for...? Check out our Retro expression vector and Lenti expression vector. Turn it on for just a little while...out Addgene's Lentiviral dCas9 SAM vector and Piggybac dCas9 SAM vector! On/off control all the time If ...Addgene's Lentiviral dTAG cloning vector and SMASh degron tagging vector! Destabilizing domains Destabilizing...resources on the Addgene blog Plasmid 101: Inducible Promoters Plasmid 101: Repressible Promoters CRISPR...Addgene's Lentiviral Tet-on and AAVS1 targeted Tet-on transgene vector. Degron tags Tagging relevant proteins...

  13. Adeno-associated virus (AAV) Plasmids

    Type
    Collection
    ...101-Viral Vector Elements Blog: AAV use in CRISPR Viral Vectors 101 eBook Packaging Plasmids AAV Plasmids... Virus (AAV) Plasmids Additional Resources: AAV Guide Biosafety Guide All Viral Plasmids Blog: Plasmids...our AAV Guide for more information about: AAV components Common uses of AAV Viral integration AAV serotypes...Helper Plasmid ID Plasmid Serotype Description PI 112867 pAdDeltaF6 All Helper plasmid for AAV packaging...Available AAV Packaging Plasmids This table contains helper plasmids for packaging your AAV product. You...table based on plasmid name, capsid, and more. ID Plasmid Gene/Insert Mutations PI AAV Plasmids This table...112862 pAAV2/1 AAV1 AAV packaging plasmid, expressing Rep2 and Cap1 Wilson 104963 pAAV2/2 AAV2 AAV packaging...

  14. Brain Initiative Collection

    Type
    Collection
    ...dependent viral vectors. Jonathan Ting 172907-AAV1 pGP-AAV-syn-FLEX-Volton2-ST-WPRE AAV-mediated expression...Addgene's AAV are made from a selected set of plasmids that are currently in our repository. AAV are distributed...Description PI 65417-AAV8 pAAV-hSyn-dF-HA-KORD-IRES-mCitrine AAV packaging vector for hSyn driven KOR ...Edward Boyden 108422-AAV5 pAAV-CAG-FLEX-Archon1-KGC-EGFP-ER2 AAV production plasmid encoding for Archon1...119742-AAV5 AAV SYN PSAM4 GlyR IRES EGFP Chemogenetics expression plasmid Scott Sternson 119744-AAV5 AAV ...element Jordane Dimidschstein 135630-AAV9 pAAV-S5E2-dTom-nlsdTom AAV vector to drive the expression of dTomato...element Jordane Dimidschstein 135631-AAV1 pAAV-S5E2-GFP-fGFP AAV vector to drive the expression of fGFP in...

  15. Serotype Testing AAV

    Type
    Collection
    ...suffix (for example, AAV1). AAV Vectors for Serotype Testing pAAV-CAG-GFP (Plasmid #37825) Description ... Viral Vector Packaging Service AAV Serotype Testing AAV Viral Vector... Vector Packaging Service: Serotype Testing AAV These AAV encode fluorescent reporters and can be used...Ready-to-use AAV in various serotypes suitable for direct injection. These AAV are made from plasmid 37825 (...Ready-to-use AAV in various serotypes suitable for direct injection. These AAV are made from plasmid 50465 (...from a vast catalog of eligible AAV plasmids and we’ll make the viral vector prep you need. Or fill out our... Add to Cart AAV Retrograde 37825-AAVrg.T 20 µL $ 140 Add to Cart pAAV-hSyn-EGFP (Plasmid #50465) Description...

  16. Caltech Systemic Capsids

    Type
    Collection
    ...Broad tropism - AAV-PHP.eB, AAV-PHP.AX, AAV-PHP.eC Cell type-biased: Astrocytes - AAV-PHP.AX with GRE ...Neurons - AAV.CAP-B10, AAV-PHP.N Broad - AAV-PHP.AX with GRE or transgenic animal L5 & Inhibitory - AAV-PHP.... Viral Vector Packaging Service AAV Caltech Systemic Capsids Viral Vector Packaging Service...from a vast catalog of eligible AAV plasmids and we’ll make the viral vector prep you need. Or fill out our...and NHPs: Broad tropism - AAV-MaCPNS2, AAV-PHP.S Sensory nervous system - AAV-MaCPNS1 Return to top...the pUCmini-iCAP-AAV.MaCPNS1 plasmid (Addgene #185136) . Browse Available MaCPNS1 AAV ID Name Promoter...the pUCmini-iCAP-AAV.MaCPNS2 plasmid (Addgene #185137) . Browse Available MaCPNS2 AAV ID Name Promoter...

  17. Retrograde AAV viral preps

    Type
    Collection
    ...Viral Vector Packaging Service AAV Retrograde AAV Viral Vector Packaging Service: Retrograde AAV As part...filtering options. Browse all Retrograde AAV *The AAV retro helper plasmid was generated as a part of the Janelia...from a vast catalog of eligible AAV plasmids and we’ll make the viral vector prep you need. Or fill out our...laboratories. These viral vector preparations were produced with the rAAV2-retro helper plasmid (Addgene #81070)...viral production , these viral vectors undergo quality control, including AAV titering by SYBR green qPCR...catalog items. Browse Available Retrograde AAV See our retrograde AAV based on the functional catagories listed...Promoter Activity Category PI 37825 AAV-CAG-GFP CAG GFP Control Boyden 51502 AAV pCAG-FLEX-EGFP-WPRE CAG EGFP...

  18. Recombinases AAV Preps

    Type
    Collection
    ...Viral Vector Packaging Service AAV Recombinases Viral Vector Packaging Service: Recombinases Cre AAV Dre...Dre AAV Flpo AAV Light-Inducible Recombinases VCre AAV Browse the following tables for in-stock AAV preps...provide high quality AAV preps from select plasmids in the repository. Browse our AAV collection of site...from a vast catalog of eligible AAV plasmids and we’ll make the viral vector prep you need. Or fill out our...See our Cre-lox Plasmids page for more information about these molecular tools. Cre AAV ID Name Promoter... rg* Wilson 24593 AAV-pgk-Cre PGK none rg* Aebischer 51507 AAV pmSyn1-EBFP-Cre Syn EBFP 5 Zeng 105540 ...50363 AAV phSyn1(S)-DreO-bGHpA Syn none 5, rg* Zeng Flpo AAV ID Name Promoter Fluorophore Serotype(s) PI ...

  19. Cre-lox system

    Type
    Collection
    ...Zebrafish Tobin 135618 pAAV-Sox2-CreERT2 Cre-ERT2 - Tamoxifen inducible; AAV donor vector AAV He 135659 pQcre1... Plasmid Collections Cre-lox Plasmids Cre-lox Plasmids Additional...Cre-lox Plasmids Addgene's Blog: Cre-lox Addgene's Blog: FLEx Vectors Cre-containing Plasmids loxP Constructs...107312 AAV-hSyn-mCherry-P2A-Cre-WPRE mCherry and Cre; expressed in neurons hSyn AAV Yang 107313 AAV-aCamkII-mCherry-P2A-Cre-WPRE-BGH-polyA...Deisseroth 55635 pAAV-EF1a-sCre SCre EF-1 alpha AAV Deisseroth 55636 pAAV-EF1a-Cre Cre EF-1 alpha AAV Deisseroth...CamKII AAV Wilson 105553 pENN.AAV.hSyn.Cre.WPRE.hGH Cre hSyn AAV Wilson 105555 pENN.AAV.hSyn.Cre.hGH Cre ...51507 AAV pmSyn1-EBFP-Cre Cre-EBFP fusion; Expression in neurons. synapsin AAV Zeng 51904 paavCAG-iCre ...

  20. Viral Production

    Type
    Collection
    ...the transfer plasmid, a plasmid encoding Rep and serotype-specific Cap genes, and a plasmid encoding adenoviral...Electron micrograph of AAV vector preparation shows that the vast majority of the vectors consist of full particles...using our virus protocols . Browse our AAV vectors collection. AAV Data Hub Lentivirus Production Lentiviral... and quality control are described below. AAV Production AAV distributed by Addgene has been produced ... with viral vector manufacturing facilities, such as the University of Pennsylvania Vector Core (Link ...Pennsylvania Vector Core. In general, titering is performed by the facility that produced the viral vector lot....determine which facility produced your viral vector lot. At Addgene, AAV particles are titered by droplet digital...

  21. Viral Prep Service for Plasmids and Pooled Libraries

    Type
    Collection
    ...technologies. Penn Vector Core AAV Vectors Many AAV vectors from the Penn Vector Core have been transferred to Addgene...eligible AAV plasmids and we’ll make the viral vector prep you need. Packaged on Request Viral Vector Packaging... Vector Packaging Service Addgene produces high-quality viral vector preps from select plasmids in our...service for plasmids and pooled libraries. Viral preparation options include lentivirus and AAV. Viral.... We know that viral vector production is a common step after requesting plasmids from Addgene, and by...In-Stock Preps We offer commonly used AAV and lentiviral vectors as in-stock preps. These high-titer preps...and ready-to-ship AAV or lentivirus prep for your next experiment today! Browse AAV | Browse Lenti Packaged...

  22. AAV Viral Preps

    Type
    Collection
    ... Viral Vector Packaging Service AAV Browse In-Stock AAV Adeno-associated virus (AAV) vectors are safe...control AAV vectors for trial purposes in serotypes AAV1, 2, 5, 8, 9 and retrograde Retrograde AAV To target...in-stock AAV preps are affordable, available in small volumes, and ready to ship. All viral vector preps ...need, try Addgene's AAV Packaged on Request service. You can choose a transfer plasmid from our repository...All Serotypes If an AAV is available in serotypes AAV1, AAV2, AAV5, AAV8, AAV9, AAVrg, the Caltech systemic...serotypes (AAV1, AAV2, AAV5, AAV8, or AAVrg). For more information, visit our Viral Vector Packaging Service...-use adeno-associated virus (AAV) available from Addgene's viral vectory packaging service. Browse constructs...

  23. PITChing MMEJ as an Alternative Route for Gene Editing

    Type
    Blog Post
    ...cloned into a vector containing Cas9 and the PITCh-gRNA. Step 3: Contransfect the PITCh vector with the vector...Guide Pages Find CRISPR Plasmids for Your Experiments Deposit Your New Plasmid Tools with Us  ...repair, making it easier to construct targeting vectors. Addgene depositor Takashi Yamamoto’s lab has harnessed...Integration into Target Chromosomes). Using their PITCh plasmids, GFP knock-in cell lines can be created in about... knock-ins, some labs have used NHEJ for whole plasmid integration; however, since NHEJ is error-prone...just upstream of a stop codon. Briefly, the PITCh vector should be constructed with 5’ and 3’ microhomology...cloning. An overview of PITCh. The PITCh plasmid contains a GFP-Puro cassette flanked by 5' and ...

  24. AAV Packaged on Request

    Type
    Collection
    ... Results Eligible Plasmids Additional Resources Adeno-associated virus (AAV) vectors are safe and useful...paper of your chosen AAV plasmid for more information on its performance. ID Plasmid Gene/Insert Promoter...Discover our AAV Packaged on Request service with all-in-one pricing for viral vector preps, which includes... Viral Vector Packaging Service Packaged on Request AAV Packaged on Request Overview...using AAV Packaged on Request, scientists can start experiments as soon as they receive their AAV prep ...transfer plasmids in our repository are eligible for this service. Select any eligible plasmid in the Addgene...transfer plasmid, including legal approval and an implementation letter DNA amplification Viral vector production...

  25. Brain Armamentarium

    Type
    Collection
    ...Gradinaru 164450-PHPeB CN1851-rAAV-hI56i-minBglobin-iCre-4X2C-WPRE3-BGHpA AAV vector for Cre expression in forebrain...CN1390-rAAV-DLX2.0-minBG-SYFP2-WPRE3-BGHpA DLX2.0 (3xCore version of DLX enhancer). AAV vector for strong... BRAIN Armamentarium Project below. Plasmids ID Plasmid Description PI Viral Preps ID Viral Prep Description...-H2B-WPRE3-BGHpA AAV vector for nuclear SYFP2 labeling in neurons. Useful for nuclear isolation and scRNA-seq...Ting Viviana Gradinaru 213945-PHPeB pAAV_BiSSTe4_ChR2_mCherry AAV construct expressing mCherry-tagged ...Fishell Viviana Gradinaru 213944-PHPeB pAAV_BiSSTe4_dTomato_nlsdTomato AAV construct expressing dTomato driven...Fishell Viviana Gradinaru 213941-PHPeB pAAV_BiPVe3_ChR2_mCherry AAV construct expressing mCherry-tagged ...

  26. University of Florida Serotype Testing Panel for the Eye and Brain

    Type
    Collection
    ...: Viral Service: All AAV Viviana Gradinaru PHP AAV Serotypes Blog: Viral Vectors As part of our Viral ...viral vector preparation. These viral vector preparations were produced with the pTR-UF11 plasmid ( Addgene...from a vast catalog of eligible AAV plasmids and we’ll make the viral vector prep you need. Or fill out our...parental AAV44.9 serotype. It was developed by Shannon Boye’s lab. Plasmid Information The plasmid below ...We provide high quality AAV preps from select plasmids in the repository. Browse the University of Florida... Viral Vector Packaging Service AAV University of Florida Serotype Testing...viral production , these viral vectors undergo quality control, including AAV titration by ddPCR, in vitro...

  27. AAV for Neuronal Tracing

    Type
    Collection
    ... Viral Vector Packaging Service AAV Monosynaptic Neuronal Tracing Viral Vector Packaging Service...from a vast catalog of eligible AAV plasmids and we’ll make the viral vector prep you need. Or fill out our...virus. AAV Vectors for Monosynaptic Neuronal Tracing ID Name Description Serotype PI 52473 pAAV-synP-FLEX-splitTVA-EGFP-B19G... Tracing These AAV encode tools for rabies virus-based monosynaptic tracing. These AAV can be used with...steps for using AAV and modified rabies virus (RABV) for monoclonal neuronal tracing. (1) AAV encoding TVA...deletion-mutant rabies virus AAV1 Wickersham 100798 pAAV-syn-FLEX-splitTVA-EGFP-tTA These AAV together can be used...Ready-to-use AAV available from Addgene's viral service encoding tools for monosynpatic retrograde neuronal...

  28. Zhang Lab CRISPR Page

    Type
    Collection
    ...Epub 2015 Apr 1. PubMed . Return to AAV In vivo plasmids AAV Plasmids with Cas9 Mouse CRISPR-Cas9 knockin... Blog Posts Jump to: SpCas9 Plasmids GeCKO SAM AAV - In vivo Delivery AAV - Cas9 Mouse CRISPR ( C lustered... lung adenocarcinoma. Delivery of a single AAV vector ( AAV-KPL ) in the lung generated loss-of-function...the mouse genome. This plasmid is used as a control for AAV-KPL. #60226 - AAV:ITR-U6-sgRNA(backbone)-pEFS-Rluc...tracrRNA: This plasmid contains three expression cassettes. To target a given site, the plasmid can be digested... in two formats: 1 vector system - lentiCRISPR - sgRNA and SpCas9 together 2 vector system - lentiCas9...of 3 vectors each available for mammalian endogenous gene activation using SAM: Addgene plasmids #61422...

  30. Chemogenetics AAV Preps

    Type
    Collection
    ...Service AAV Chemogenetics Viral Vector Packaging Service: Chemogenetics AAV Use our chemogenetics AAV to chemically...from a vast catalog of eligible AAV plasmids and we’ll make the viral vector prep you need. Or fill out our...filtering options. *AAVrg = retrograde serotype, produced with the AAV retro helper plasmid from the Karpova...119742 AAV SYN PSAM4 GlyR IRES EGFP PSAM4 GlyR - Inhibition IRES EGFP none 5 Sternson 119744 AAV CAMKII... labs. See the Retrograde AAV Inventory Page . Browse all Chemogenetics AAV Don’t See What You’re Looking...Constitutive (non-cre dependent) Serotype AAV1 AAV2 AAV5 AAV8 AAV9 Retrograde AAV PHPeB Clear Filters ID Name Receptor...-to-use AAV available from Addgene's viral service encoding chemogenetics tools. Viral...
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