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  1. Lentivirus Production

    Type
    Protocol
    ...determine what ratio gives the highest percentage of GFP positive cells. Refer to the table below for a possible...Data Figure 1: 293T cells were transfected with the GFP-expression plasmid pRosetta using μg total DNA to...efficiencies as measured by the highest proportion of GFP positive cells without limiting cell growth. Left...Left panels: bright field images; right panels: GFP channel images. Last reviewed on: August 2, 2023... Protocols Lentivirus Production Lentivirus Production You may also ...This protocol can be used to produce lentivirus from a lentiviral vector transfected into 293T cells using...this protocol to generate lentivirus Protocols...
  2. General Transfection

    Type
    Protocol
    ...cells optimized for viral production (AAVpro or Lenti-X), but any HEK293T cell line should work. This ...determine what ratio gives the highest percentage of GFP positive cells. Refer to the table below for a possible...checking for protein expression. Sample Data Legend: Lenti-X 293T cells were transfected using 1:1, 1:2, 1:...
  3. Quick Guide to All Things Lentivirus

    Type
    Blog Post
    ...containing the gene of interest (pTet-IRES-EGFP, pPRIME-Tet-GFP-FF3) and one with either tTA or rtTA (FUW-M2rtTA...viral titre to the number of cells transduced by a GFP encoded virus, and the reverse transcriptase activity...property makes the lentivirus an attractive choice for a gene transfer vector. Lentiviral vectors Although...generation of lentiviral vectors consisted of three plasmids encoding 1) the lentiviral vector genome ...used. Inducible lentiviral transfer vectors The most widely used inducible lentiviral vector system is...Your Research Lentiviral Vector Uses and Overview Resources on Addgene.org Find Lentiviral Vectors for...Ready-to-Use Lentivirus Browse Our Lentivirus Guide Pages  ...
  4. Lentiviral Prep Service

    Type
    Collection
    ...Purpose PI 17446 pLenti CMV GFP Hygro (656-4) GFP Hygromycin 3rd gen lentiviral eGFP expression vector...dCAS9-VP64_GFP dCAS9 (D10A, H840A) none Expresses dCAS9-VP64 activator with 2A GFP Zhang 61425 lenti dCAS-VP64... dCAS9 (D10A, N863A) Blasticidin 3rd generation lenti vector encoding dCAS9-VP64 with 2A Blast resistance... Viral Vector Packaging Service Lentivirus Browse In-Stock Lentivirus Pooled ...sgRNA library Brunello in lentiGuide-Puro Human sgRNA library in backbone lentiGuide-Puro targeting 19,114...gRNA pooled library in lentiCRISPRv2 Human sgRNA library in backbone lentiCRISPRv2 targeting 19,114 genes...sgRNA library Brie in lentiGuide-Puro Mouse sgRNA library in backbone lentiGuide-Puro containing 78,637...
  5. Lentivirus Plasmids

    Type
    Collection
    ...hUbC-driven EGFP; can be used for cDNA expression Baltimore 12247 pLVTHM 2nd EF-1a-driven GFP and shRNA ... a reporter. See plasmid 17618 for GFP plasmid. Cheng 17452 pLenti CMV Puro DEST 3rd Gateway destination...gRNA plasmids table and search "lenti". ID Plasmid Generation Description PI 8453 pLKO.1 puro 3rd U6-driven...Blog: Tips for Titering Your Lentiviral Preps Lentiviral Pooled Libraries All Viral Plasmids Packaging Plasmids...researcher safety. Read our Lentiviral Guide for more information about: Lentiviral components 2nd versus 3rd...generation lentiviral systems Viral production Frequently asked questions about lentivirus use Glossary...Glossary of commonly used lentiviral terms Popular Lentiviral Envelope and Packaging Plasmids One safety feature...
  6. Empty Backbones - Choosing Your Perfect Plasmid Backbone

    Type
    Collection
    ...expression vector GFP Varies pMKO.1 GFP - Retroviral shRNA expression pLenti CMV GFP DEST - ...Fluorescent proteins (GFP, mCherry, etc) Localization pcDNA3-EGFP - C-terminal GFP for mammalian expression...genome MSCV-IRES-GFP or pMSCV-IRES-mCherry FP - Mammalian retroviral gene expression with GFP or mCherry selectable... Mammalian lenti sgRNA(MS2)_zeo backbone - 3rd gen lenti sgRNA cloning backbone with MS2 loops at tetraloop... genome pLenti CMV GFP DEST - Lentiviral Gateway destination vector for gene expression pLenti CMV/TO ...-zeo resistance marker pLenti CMV/TO GFP-Zeo DEST (719-1) - 3rd gen lentiviral Gateway destination vector...CMV/TO promoter, GFP-Zeo pLenti CMV/TO Zeo DEST - Tet-inducible lentiviral Gateway destination...
  7. Tips for a 1st Time CRISPR User (by a 1st Time CRISPR User)

    Type
    Blog Post
    ...from BRAF-edited and GFP-edited amplification products (BRAF-GFP duplex) and from GFP-edited amplification... for gene editing. A virus encoding GFP plus a gRNA targeting GFP (pXPR_011, 59702-LV) was used to assay... by the virally-delivered GFP gRNA, will edit out the virally-delivered GFP gene. So, cells with no Cas9...I thought I could see GFP expression with a fluorescence microscope, but the GFP expression was so low...expressed GFP) that I couldn’t get a good image. You could probably also do Western blotting against GFP expression...amplification products (GFP-GFP duplex) as a control. 4. After optimizing the reaction and confirming the presence... didn’t. 2. In the original publication they see GFP expression in about 20% of their Cas9-expressing ...
  8. Hot Plasmids - January 2023

    Type
    Blog Post
    ...released Anti-GFP [N86/8R] as another option for targeting GFP. But why might you need a second GFP antibody...PMCID: PMC9015817.   Meet anti-GFP [N86/8R]! by: Ashley Waldron Anti-GFP [N86/38.1R] was among the first... targeting system Heat-inducible lentiviral Car constructs Anti-GFP [N86/8R] ready-to-use antibody Hyperfolder...western blot for mEGFP-HRas (from Plasmid 18662). See the Applications section on the Anti-GFP [N86/8R] page...?  Similarities:  Generated against full length GFP and converted to recombinant form by the NeuroMab...variable domains that recognize different epitopes on GFP Monoclonal antibodies can yield relatively low signal... details. Image credit: Addgene.   Find anti-GFP [N86/8R] here!   Andrews NP,  et. al. A toolbox of...
  9. New and Upcoming Viral Vectors - May 2020

    Type
    Blog Post
    ...respectively of a GFP dependent Cre. The two segments come together in the presence of GFP to form a functional... activation to cells expressing both Cre and GFP. The GFP dependent FLPo, Flp-DOG (75469), is also available...targeted expression DREADDs GFP dependent Cre and FLPo And more!  New lentiviral preps SARS-CoV-2 protein...in new serotypes, as well as a new Cre dependent GFP control plasmid for expression in forebrain GABA-...Karl Deisseroth 83895 AAVrg pAAV-hDlx-Flex-GFP-Fishell_6 Gordon Fishell   Optogenetics Optogenetics...transgenic reporter lines that supply a variety of GFP expression patterns, thus allowing you to further...AAV2 pAAV-Ef1a-fDIO mCherry 83900 AAVrg pAAV-mDlx-GFP-Fishell-1 Biosensors Plasmid Serotype Name ...
  10. Hot Plasmids: Spring 2025

    Type
    Blog Post
    ...Addgene's own Lentiviral backbone plasmids here! Figure 2: Addgene cloned EGFP into pAG Lenti CMV N-HA...#236079) to create pAG Lenti CMV HA-EGFP Puro (Addgene #236081), then generated lentiviral particles in HEK293T...and labeled with primary antibody (0.01 µg/mL Anti-GFP [N86/38.1R] (Addgene #180084), 0.01 µg/mL Anti-HA...share our very own set of empty, HA-, and EGFP-expressing lentiviral backbones designed right here at Addgene...tailored deactivation rates Addgene's own empty lentiviral backbones New recombinant antibodies for Glypicans... packaging and strong expression after inserting EGFP and provide step-by-step protocols for viral generation...Figure 2). Looking for more? Explore our empty and EGFP retroviral plasmid set, our bacterial plasmid set...
  11. 15 Years of Addgene: The Top 15 Plasmids

    Type
    Blog Post
    ...The 2A-GFP allows for screening of transfected cells. Find pSpCas9(BB)-2A-GFP (PX458). lentiCRISPR v2 -...lines already expressing Cas9. Find lentiGuide-Puro. pSpCas9n(BB)-2A-GFP (PX461) - Similar to pSpCas9(BB)...requested plasmid of 2017. Find pMD2.G. pSpCas9(BB)-2A-GFP (PX458) - This plasmid from the Zhang lab expresses...BB)-2A-Puro (PX458), pSpCas9n(BB)-2A-GFP (PX461) expresses both a Cas9 protein and a gRNA. But instead ...nick, rather than cleave DNA. Find pSpCas9n(BB)-2A-GFP (PX461). pSpCas9(BB)-2A-Puro (PX459) - This mammalian...promoter. It’s a 3rd generation lentiviral backbone that must be used with lentiCas9-Blast or with cell lines...the last 15 years - the majority of these being lentiviral or CRISPR tools. How many times have these plasmids...
  12. X-CHIME: Context Dependent Germline Knockout in Immune Cells

    Type
    Blog Post
    ... out a pair of genes pXPR_219 Violet-excited GFP (vex) Rosa26-Cas9 I-CHIME Inducible knockout...knockout of a single gene pXPR_068 Violet-excited GFP (vex) Rosa26-FlpO-ERT2; H11-Cas9 L-CHIME ...types (lineage-based) pXPR_070 Violet-excited GFP (vex) Lineage-Cre; Rosa26-Cas9 S-CHIME Knocks...of genes sequentially pXPR_220 Violet-excited GFP (vex) Rosa26-FlpO-ERT2; H11-Cas9 Table 1: X-CHIME...(Rosa26-Cas9) hematopoietic stem cells with a lentiviral vector containing a gRNA. These transduced stem...systems which utilizes custom mouse strains and lentiviral expression vectors (Table 1). The four systems...pXPR_071, a plasmid that leads to higher titer lentivirus preparations than the first generation CHIME ...
  13. 22 Hot Plasmid Technologies from 2014

    Type
    Blog Post
    ...dim GFPs The availability of the three dimensional structure of GFP, its brighter mutant S65T-GFP, and...and GFP shRNA. Mice carrying the R26DsRedR; CRUSH-GFP; and Nestin-Cre alleles showed efficient GFP knockdown...identified a family of GFP proteins in the cephalochordate Branchiostoma floridae, named bfloGFPs. They went on...and demonstrate that they can successfully create GFP fusion proteins with a variety of genes across the...enabled scientists to engineer a wide variety of GFPs with diversity in fluorescence brightness, intensity...Cas9 gene editing. Dr. Masahito Ikawa has created a GFP reporter plasmid for scientists to validate the efficacy...plasmid undergoes HR or SSA to reconstitute functional GFP, and the cells will turn green. Mashiko et al.,...
  14. Popular Retroviral Vectors and Their Uses in Scientific Research

    Type
    Blog Post
    ...technology CAG-GFP-IRES-CRE (Plasmid #48201) This retroviral vector from the Gage lab encodes GFP and Cre-recombinase...Vectors   Expressing your gene of interest pMKO.1 GFP (Plasmid #10676) Retroviral vector for cloning and...and expressing your gene of interest with GFP expression.  Cloning and expressing RNAi pMKO.1 puro (Plasmid...descriptions of the lentiviral elements and the 2nd and 3rd Generation Lentiviral Systems can be found...found in our Lentiviral guide. Popular packaging and lentiviral envelope plasmids are listed here. Please...pre-treated with a compound of interest. Lentiviral vectors Popular Lentiviral Transfer Vectors    Expressing your...3rd generation lentiviral vector backbone from the Shih lab. You can find more lentiviral expression vectors...
  15. Kazuhiro Oka Lentiviral Vectors

    Type
    Collection
    ...Vectors Lentivirus Oka Lab Lentiviral Vectors Baylor College of Medicine Gene Vector Core Lentiviral and ...promoter pCDH-EF1-copGFP 73030 Expresses copGFP from the EF1 promoter pCDH-EF1s-Nluc-P2A-copGFP-T2A-Puro 73032...pCDH-EF1s-copGFP 73034 Expressed copGFP from a truncated EF1 promoter pCDH-CMV-Nluc-P2A-copGFp-T2A-Puro...Resources Addgene Blog: Your Lentiviral Plasmid FAQs Biosafety Resource Guide Lentiviral Transfer Vectors Adenoviral...deposited a series of lentiviral and adenoviral transfer vectors with Addgene. Lentiviral Transfer Vectors ...infectious virus. Protocol: Lentivirus Transfer Vector Production Protocol Lentiviral Vector ID Purpose pCDH-EF1...pCDH-EF1-DIO-copGFP 72253 Expresses copGFP under EF-1 promoter when Cre is expressed by another vector pCDH-EF1...
  16. Multiplex Genome Editing with CRISPR-Cpf1

    Type
    Blog Post
    ...array plus a GFP tag. Four weeks post-infection, there was strong expression of Cpf1 and GFP in the targeted...4 targets when an array of 4 crRNAs, Cpf1, and a GFP tag were transfected on a single plasmid. This is...~75% of neurons were co-transduced with Cpf1 and GFP.  Of these neurons, ~15% had indels at all 3 targeted...Multiplexed Lentiviral Expression Cassettes lentiviral transduction One vector expresses Cas9, eGFP, and ...expressing the Cpf1 multiplex system from a lentiviral vector.  Lentiviruses carry a (+) strand RNA copy of the... options for Cpf1 multiplexing: Transfection, lentivirus, and AAV Cpf1’s ability to process its own pre-crRNA... multiple expression platforms: transfection, lentiviral transduction, and AAV transduction.  In most ...
  17. Viral Production at Addgene

    Type
    Blog Post
    ... later, Cre-dependent GFP expression was detected with direct fluorescence. GFP was not detected in the...viral genomes (vg)/cell, pAAV-CAG-FLEX-rc [Jaws-KGC-GFP-ER2] (Addgene 84445-AAVrg) alone at 1.1E6 vg/mL, ...prep # 55632-AAVrg). pAAV-CAG- FLEX-rc [Jaws-KGC-GFP-ER2] was a gift from Edward Boyden (Addgene viral...Browse our AAV vector collection. Lentivirus Production Lentiviral preparations ("-LV” catalog items)...Concentrated lentiviral preparations (“-LVC” catalog items) are generated from lentiviral preparations...being frozen. Specifically, lentiviral particles are collected from the lentiviral preparation by precipitation...for in our reported titers. Lentiviral vectors are titered using a lentiviral droplet digital PCR (ddPCR...
  18. CRISPR Plasmids - Empty gRNA Vectors

    Type
    Collection
    ... pCAG-SpCas9-GFP-U6-gRNA 79144 Mammalian hU6 yes, cut S. pyogenes EGFP Zou pCAG-eCas9-GFP-U6-gRNA 79145...S. pyogenes Puro Zhang lenti sgRNA(MS2)_zeo backbone 61427 Mammalian/Lentiviral BsmBI none S. pyogenes...Mammalian/Lentiviral LIC none S. pyogenes GFP Vakoc U6>sgRNA(F+E) 59986 Other/Ciona intestinalis BsaI none S....Yang pLCKO 73311 Mammalian/Lentiviral hU6 none S. pyogenes Puro Moffat lenti sgRNA(MS2)_puro backbone 73795...pyogenes URA3 Davis lenti sgRNA(MS2)_puro optimized backbone 73797 Mammalian/Lentiviral U6 none S. pyogenes...cut Lb Cpf1 Neo Welker AIO-GFP 74119 Mammalian U6x2 yes, nick S. pyogenes EGFP Jackson AIO-mCherry 74120...Mammalian/Lentiviral BbsI none S. pyogenes Puro, TagBFP Yusa lentiGuide-Puro 52963 Mammalian/Lentiviral BsmBI...
  19. Hot Plasmids - June 2020 - Barcoded CRISPR Library, Sparse Cell Labeling, Calcineurin Reporter, and DNA Staining Dye Alternative

    Type
    Blog Post
    .... https://doi.org/10.1101/788679 A destabilized GFP reporter for calcineurin activity in yeast Article...reporter consisting of a quickly-degraded destabilized GFP variant fused to a 4X CDRE repeat. In addition to... Ca2+ (bottom panels) equipped with destabilized GFP calcineurin reporter (blue) and stained with PI (...death. Image from Diessl et al., 2020. Find the GFP reporter calcineurin activity here Diessl et al.,...infecting cells with sgRNAiBAR library containing lentiviral vectors the iBARs can be detected by next generation... They created plasmids for purification of tTALE-eGFP and tTALE-mCherry. In comparison to intercalating...
  20. 28 Hot Plasmid Technologies from 2015

    Type
    Blog Post
    ...both the GFP and the RFP proteins in the nucleus. During the first round of translation, the GFP proteins...Compartment mCherry-Rab5 BFP-Rab5 GFP-Rab5B Vacuolar & Budding Compartment GFP-Rab4B Mitochondria mito-BFP...GTPase in division) GFP-Mff (outer membrane protein) Late Endosome mCherry-Rab7A GFP-Rab7A     ...protein Knock-off, is made up of three components: a GFP protein that binds to the coding region of a reporter...reporter mRNA via a PP7 coat protein (NLS-PCP-GFP) a RFP protein that binds to the 3’ UTR of the reporter...these dCas9s with one of three fluorescent proteins (GFP, BFP and mCherry), the authors were able to visualize...monoclonal antibodies against them, fused them to GFP, and compared them to popularly used epitopes tags...
  21. Hot Plasmids June 2018 - Reverse transcriptase, nanobody, and protein-DNA interaction tools

    Type
    Blog Post
    ...includes a set of plasmids each containing an anti-GFP nanobody fused with a different functional element...using different detection methods. Since the anti-GFP nanobody is able to bind fluorescent proteins derived...here can be used to target any protein fused with GFP, YFP, mCerulean, or other related fluorophores. Find...specific DNA sequence in only 1 min. Using these lentiviral plasmids, catalytically inactive Cas9 (dSpyCas9...
  22. CRISPR 101: Mammalian Expression Systems and Delivery Methods

    Type
    Blog Post
    ... create a fusion with a reporter protein such as GFP? All of these outcomes can be most effectively achieved...typically used for gRNA. May contain reporter gene (e.g. GFP) to identify and enrich positive cells or a selection...transfer vectors. May contain reporter gene (e.g. GFP) to identify and enrich positive cells. Likely contains...efficiency. Lentiviral transduction Cas9 and gRNA can be present in a single lentiviral transfer vector...plasmids provide the necessary components to make lentiviral particles. Stable, tunable expression of Cas9...Difficulty Very easy Very hard Viral delivery   Lentivirus Retrovirus AAV Cells Dividing or nondividing...: Traffic Light Reporter System: Can evaluate lentiviral component delivery as well as genome repair by...
  23. Hot Plasmids and Viral Preps - September 2021

    Type
    Blog Post
    ...coding sequence for a green fluorescent protein (GFP). The success of the A-to-G conversion could be seen...through the success of the change in expression of GFP, for the stop codon should have changed from “TGA...cytometry is used to sort populations by mCherry and GFP expression to check for ABE’s. b) Quantification ... 10 guide RNAs per gene. The lab modified the lentiviral backbone to encode the CD90.1 selection marker...
  24. Hot Plasmids - November 2023

    Type
    Blog Post
    ...transcriptional activation events as bands of induced GFP-iPAK4 expressed under promoters like the tet-responsive...fluorescent signal in the ticker tape from cFos::GFP-iPAK4 within an hour. Analyzing the ticker tape images...to red at 12 h, and PMA activation induces cFos::GFP-iPAK4 at 3 or 6 h. Scale bar 5 µm. Images adapted...components to reprogram delivery vehicles like lentiviral vectors, CreVLPs, and eVLPs for in vivo and in... to target multiple delivery systems, such as lentivirus and MMLV gag, to specific cell types. It can ...cytoplasm and can be fluorescently labeled. Using lentiviral vectors to stably or inducibly express tagged...
  25. Neurodegeneration Plasmid Collection

    Type
    Collection
    ... PINK1 N-GFP PINK1 GFP CMV Parkinson's Mark Cookson 13316 pcDNA-DEST47 PINK1 C-GFP PINK1 GFP CMV Parkinson's...21190 GFP-pcDNA3-PKCgamma-cys1Acys1B PRKCG GFP CMV Spinocerebellar ataxia 27 Tobias Meyer 21204 GFP-N2-PKCgamma...PKCgamma PRKCG GFP CMV Spinocerebellar ataxia 26 Tobias Meyer 21205 GFP-C1-PKCgamma-C1A PRKCG GFP CMV Spinocerebellar...-HtrA2-EGFP HTRA2 GFP CMV Parkinson's L. Miguel Martins 14122 pcDNA3-HtrA2-EGFP S306A HTRA2 GFP CMV Parkinson's... GPD 25QDProGFP p416 HTT GFP GPD Huntington's Susan Lindquist 15569 GPD 104QDProGFP p416 HTT GFP GPD Huntington's...GAL 25Q+ProGFPp416 HTT GFP GAL1 Huntington's Susan Lindquist 15581 GAL 46Q+ProGFPp416 HTT GFP GAL1 Huntington's...GAL 72Q+ProGFPp416 HTT GFP GAL1 Huntington's Susan Lindquist 15583 GAL 103Q+ProGFPp416 HTT GFP GAL1 Huntington's...
  26. Viral Vectors 101: Biosensors

    Type
    Blog Post
    ...interest (analyte), and accumulation of ATP activated GFP expression (receptor), then the transducer could ...plate reader, or any other instrument which measured GFP levels and converted that signal into a readable ...ready to deliver your sensor, whether you choose lentivirus, gamma retrovirus, or AAV as your vehicle, we...
  27. Luciferase Plasmid Collection

    Type
    Collection
    ...plasmid Attila Karsi 105621 Lenti-luciferase-P2A-Neo Firefly EF1α Lentiviral expression of firefly luciferase...Root 105533 pAAV.CMV.Luc.IRES.EGFP.SV40 Firefly CMV AAV expression of firefly luciferase and GFP James Wilson...phGluc Gaussia EF1α Expression of Gaussia luciferase; GFP is expressed if cells are infected with virus Christopher...Firefly TGB AAV expression of firefly luciferase and GFP James Wilson 106457 pCR3.1-Luc Firefly CMV Mammalian...fusions using Gateway cloning. Lentival. Mikko Taipale 87078 pLenti6.2-Nanoluc-ccdB NanoLuc® Creation ...fusions using Gateway cloning. Lentiviral. Mikko Taipale 87077 pLenti6.2-Firefly-ccdB Firefly Creation ...transfection Feng Zhang 21471 pLenti PGK V5-LUC Neo (w623-2) Firefly PGK Lentiviral expression of firefly luciferase...
  28. Hot Plasmids Spring 2024

    Type
    Blog Post
    ...Figure 5: A) Representative confocal images showing GFP expression from ssAAV5s with hIBA1 or hIBA1a promoters...Quantifications showing high microglia specificity of GFP expression for the indicated ssAAV5s. C) Quantifications...within the lentiviral 3’ long terminal repeats (LTR). The 3’ LTR is duplicated during lentiviral integration...license. In this iteration, the size of the lentiviral insert was minimized to reduce recombination ...III transcript, the sgRNAs were encoded on the lentiviral minus strand in the 3’ LTR. These optimizations...
  29. SARS-CoV-2 Pseudotyped Virus

    Type
    Collection
    ...expressing destabilized EGFP. Lenti-luciferase-P2A-Neo - Lentiviral reporter vector expressing firefly luciferase...firefly luciferase. pLentiEGFPdestablized - EFS-EGFPd2PEST-2A-MCS-Hygro - Lentiviral vector expressing ...examples of each in Addgene's collection: HIV-based lentiviral particles Murine leukemia-based retroviral particles...Plasmids Envelope and Packaging Plasmids Popular Lentiviral Envelope and Packaging Plasmids Popular γ-Retrovirus...neutralizing activity are listed below: pBOBI-FLuc - Lentiviral reporter vector expressing firefly luciferase...firefly luciferase. pHAGE-CMV-Luc2-IRES-ZsGreen-W - Lentiviral dual reporter vector expressing firefly luciferase...luciferase and ZsGreen. NL4-3 mCherry Luciferase - Lentiviral dual reporter vector expressing mCherry and firefly...
  30. Using Addgene's New Viral Service for Your Research

    Type
    Blog Post
    ... CRISPR and so we also have lentivirus for delivering control shRNAs, GFP, and rtTA from the Tet-on system... Service Suggestion Form. Lentivirus As some of the most popular lentiviral vectors in the repository ... of our new Viral Service, we’re distributing lentivirus (with many CRISPR tools included among the preps...collection. We’ll provide a brief rundown of what lentiviral and AAV preps are available below, but you can... tools, we’re starting the Viral Service with lentiviral preps for the delivery of Cas9 endonuclease, ...we’ve got viruses to help you out. Of course, lentiviruses are used in experiments other than just those...with viruses including advice on how to titer lentiviral preps, troubleshoot viral transductions, and ...
  31. Cre-lox system

    Type
    Collection
    ...CAG-Cremyc-2A-GFP GFP and Cre CAG Mammalian Lu 117148 Hiv7CMV-Cremyc-2A-GFP GFP and Cre CMV Lentiviral Lu 118029...48201 CAG-GFP-IRES-CRE Cre and GFP coexpression CAG Retroviral Gage 49054 CAG-GFP/cre Cre-GFP fusion CAG...iRFP670-EFS:Cre-2A-GFP iRFP670, Cre, and GFP TRE Mammalian Jacks 68544 AAV-Cre-GFP Cre CMV AAV Nestler...13770 pCALNL-GFP Cre dependent GFP expression Mammalian Cepko 8389 p212 pCMV-EGFP/RFP EGFP-dsRed gene switch...promoterless CRE-GFP fusion none Mammalian Sauer 11960 pBS537 tet-hCMV-GFPcre tet inducible Cre-GFP fusion tet-hCMV... shRNA coexpression CMV Lentiviral Fuchs 26646 pCAG-Cre-IRES2-GFP Cre and GFP coexpression CAG Mammalian...CAG Retroviral Gage 49056 AAV-GFP/Cre Cre-GFP fusion CMV AAV Gage 49111 pEMS1980 iCre with MCS for inserting...
  32. New Optogenetic Tools for Cytoskeleton and Membrane Control

    Type
    Blog Post
    ...activity can be monitored by phosphatidic acid probe GFP-PASS. B) Time series of HEK 293T cells expressing...membrane-targeted SuperPLDx30 (magenta) and PA probe GFP-PASS (green). After 30 minutes of blue light illumination...SspB-mCherry-p60 katanin module, which are also available as lentiviral plasmids if you want to make a stable cell line...
  33. Fluorescent Protein Guide: Subcellular Localization

    Type
    Collection
    ... Gerlich 25999 LV-GFP Chromatin H2B GFP Elaine Fuchs 11680 H2B-GFP Chromatin H2B GFP Geoff Wahl 21210 ...61803 GFP-Rab7A Late endosomes Rab7a AcGFP Gia Voeltz 12605 GFP-rab7 WT Late endosomes RAB7 GFP Richard...89472 GFP-hChibby1 Mother Centrioles / Ciliary Base Chibby1 GFP Ken-Ichi Takemaru 89770 pLenti-EGFP-hChibby1...Gadella 50057 pLYS1-FLAG-MitoGFP-HA Mitochondria MCU GFP Vamsi Mootha 49153 GFP-Mff Mitochondria-Outer ...Filaments beta-actin EGFP Robert Singer 27382 pDEST/N1-hEB1-GFP Microtubules EB1 GFP Robin Shaw 40908 pDEST...Beta-catenin GFP Adherens Junctions Beta-catenin EGFP Alpha Yap 67937 mouse E-cadherin GFP (423) Adherens...38277 pMXs-puro GFP-p62 Autophagosome Sequestosome-1 EGFP Noboru Mizushima 38272 pMXs-IP GFP-WIPI-1 Autophagosome...
  34. Adenovirus Plasmids

    Type
    Collection
    ... in E3 and insertion of GFP expression cassette. Bunz 179202 pAd5-B6/7 ∆E3-GFP Adenoviral A block for ...regions present in pAd5-B6 ∆E3-GFP and pAd5-B7 . Bunz 179203 pAd5-B1-deltaE1-GFP Adenoviral A block for the...production of GFP-trackable viruses containing transgene under CMV promoter Vogelstein 16407 pAdEasy 2-GFP beta-gal...with large inserts Vogelstein 179201 pAd5-B6 ∆E3-GFP Adenoviral A block for the AdenoBuilder genome assembly...system. Block 1 with deletion in E1 and insertion of GFP expression cassette. Bunz 16402 pShuttle Shuttle ...Vogelstein 16404 pAdTrack Shuttle For production of GFP-trackable viruses containing transgene under a chosen...beta-gal Shuttle Test plasmid that contains β-gal and GFP; contains ∼10Kb genomic DNA stuffer Vogelstein 18703...
  35. Multiplexed Overexpression of Regulatory Factors (MORF) collection

    Type
    Blog Post
    ...library comes either with or without mCherry and GFP fluorescent controls. Alternatively, you can order...Blog Learn More about Lentivirus Figure out How to Get the Most from Your Lentiviral Transduction Additional... individually overexpressed and available for lentiviral packaging and delivery to target cells as a pooled... Fig. 1: Barcoded TF library was packaged in lentiviral particles and used to transduce human embryonic...
  36. CRISPR 101: Multiplex Expression of gRNAs

    Type
    Blog Post
    ...destination vector contains GFP, enabling you to select cells with high GFP expression. These cells have...Multiple Lentiviral Expression Systems (MuLE) Kit: This kit can be used to create lentiviral vectors expressing...CRISPR/Cas9-based genome engineering from a single lentiviral vector.” Nucleic Acids Research 42(19) (2014)...
  37. Tetracycline Inducible Expression

    Type
    Collection
    ...option rtTA On Kowarz 16623 pBI-GFP Expression of your gene of interest & GFP from a bidirectional tet-responsive...Either Lung 11662 pPRIME-TET-GFP-FF3 Lentiviral, miRNA expression (PRIME) system for application in knockdown...Locus. Tet-On 3G On Doyon 58245 pGLTR-X-GFP Single vector lentiviral Gateway RNAi system for conditional ...cells; Expresses firefly luciferase hairpin and GFP under pTREtight promoter None Either Elledge 35625...generation; contains expression cassette for TetR-P2A-GFP; see article for additional constructs TetR On Geley...inducible promoter at its 3' end, which controls GFP expression tTA Off Verma 26803 pEnt L1L3 EF1a-tTA...pTet-IRES-EGFP Lentiviral plasmid for inducible expression of transgene of interest and EGFP None Either...
  38. Choosing Your Perfect Empty Backbone

    Type
    Blog Post
    ...using a reporter vector to tag YGOI with lacZ and/or GFP (e.g. pPD80_08) and following the expression pattern... allow you to visualize YGOI in vivo (try pcDNA3-EGFP). If you can get away with generating transiently...basic mammalian expression system and go for a lentiviral or a retroviral backbone. This is considered ...
  39. Fluorescent Protein Guide: Empty Backbones

    Type
    Collection
    ...Expression pRSETA-PAGFP - Bacterial Expression tandem PA-GFP-KanR - Yeast Expression pFA6a-PA-GFP-kanMX6 - Yeast...Expression pFA6a-link-yEPAGFP-CaUra3 - Yeast Expression mPA-GFP-N1 - Mammalian Expression mPA-GFP-C1 - Mammalian...then scientists have engineered numerous GFP-variants and non-GFP proteins that result in a diverse set ...tagging with mCherry, mOrange, mCerulean pET GFP - C-terminal GFP for bacterial expression Davidson Lab Plasmids...Expression EGFP 488 507 34 6 Prone to dimerization pcDNA3-EGFP - Mammalian Expression pCAG-GFP - Mammalian...Mammalian Expression PA-GFP 504 517 14 Monomer (A206K) pPAGFP-N1 - Mammalian Expression pPAGFP-C1 - Mammalian ...with mCherry, Cerulean, Citrine, and eGFP pPD95_75 - C-terminal GFP for C. elegans expression pHT101-mCherry...
  40. Viral Vectors 101: Virus Safety

    Type
    Blog Post
    ... for a researcher to come into contact with than GFP. Similarly, if the viral vector carries an shRNA ...viruses grown in the lab are gamma retrovirus, lentivirus, adenovirus, and adeno-associated virus (AAV)...humans, earning this virus a BSL-2 classification. Lentivirus (a type of retrovirus) and gamma retrovirus require...and required in BSL-2 labs. When working with lentivirus, many organizations require or recommend disposable...you have to work with BSL-2 materials, such as lentivirus. Most of this work will need to be done in a ...that particles are not sprayed into the air. For lentivirus and other BSL-2 materials, you’ll need to use...for extra security. Finally, when spinning down lentivirus, you’ll need to use centrifuge bin covers, which...
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