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We narrowed to 111 results for: lentiviral dna vectors

Showing: 41 - 60 of 111 results

  1. New Tool for Lineage Tracing: The ClonTracer Library

    Type
    Blog Post
    ...consists of a pool of lentiviral vectors with roughly 73 million semi-random, 30 bp DNA barcodes that are ...individually label millions of mammalian cells through lentiviral infection and to monitor their abundance and ...optimize protocols for in vivo screening with lentiviral pooled shRNA libraries. Later, Bhang says, “When... Find the ClonTracer Library at Addgene Find Lentiviral Plasmids at Addgene Check out Our Cancer Resources...infection. When using the ClonTracer system, lentiviruses containing these barcodes infect the target ...

  2. Hot Plasmids - June 2020 - Barcoded CRISPR Library, Sparse Cell Labeling, Calcineurin Reporter, and DNA Staining Dye Alternative

    Type
    Blog Post
    ...infecting cells with sgRNAiBAR library containing lentiviral vectors the iBARs can be detected by next generation... as a DNA staining dye Article contributed by Jennifer Tsang Figure 4: Double stranded DNA bound... fusions with tTALE to “stain” DNA. TALE proteins can bind specific DNA sequences which brings the attached...4X CDRE repeat. In addition to showing that this vector is suitable for flow cytometry and fluorescence...alternative to intercalating fluorescent dyes for DNA-protein interaction studies? Kyubong Jo's lab generated...attached fluorescent protein to the DNA. They created plasmids for purification of tTALE-eGFP and tTALE-mCherry...intercalating fluorescent dyes which can cause photo-induced DNA breakage and structural deformation, these probes...

  3. New Tools Enable CRISPRa for Neuroscience Applications

    Type
    Blog Post
    ...the Day Lab, who developed neuron-optimized lentiviral vectors for in vitro and in vivo CRISPRa (Savell ...physiology. The Day Lab’s neuron-optimized lentiviral vectors can be found here. High-throughput screening...Neuron-optimized viral vectors Alternatively, Cas9 activators can be delivered to the CNS by lentivirus, which has.... This dual lentivirus system pairs ubiquitous expression of sgRNA(s) and mCherry (vector 1) with neuron-specific...their optimized vectors to highlight a key advantage of CRISPRa over traditional cDNA overexpression: ...including transgenic mice, neuron-optimized viral vectors, and high-throughput screening approaches. Here...largely precludes delivery by adeno-associated vectors (AAV). This issue was recently circumvented by ...

  4. 5 Tips for Troubleshooting Viral Transductions

    Type
    Blog Post
    .... Viral vectors undergo DNA rearrangements. Depending on the composition of the viral vector and the ... about Lentiviral Vectors Additional Resources on the Addgene.org Check out Our Viral Vector Guide Pages...hepatitis B virus DNA and hepatitis C virus RNA quantification as measured with branched-DNA technology." ... one point or another. After amplifying a viral vector, collecting virus, and transducing target cells...the repeat sequences present in the LTR, the vector may undergo genomic rearrangements during replication...genomic rearrangements, try amplifying the viral vector with bacteria designed to minimize rearrangements...have low titers (which is unavoidable for some vectors) you can concentrate your virus by ultracentrifugation...

  5. Lentivirus Production

    Type
    Protocol
    ...from a lentiviral vector transfected into 293T cells using a polyethylenimine (PEI) transfection protocol...polyethersulfone (PES) filter Transfection grade DNAs: Lentiviral Packaging plasmid(s), Envelope plasmid, and...plasmid DNA purification should include an endotoxin removal step. For high quality plasmid DNA, the plasmids...the diluted DNA mixture. Add the diluted PEI dropwise while gently flicking the diluted DNA tube. Incubate...Viral Vector Guides Mol Bio Protocols Introduction This protocol can be used to produce lentivirus from...possible range of ratios to test: Ratio of DNA:PEI μg of DNA μL of 1 mg/mL PEI 1:1 18.9 18.9 1:2 18.9 37.8...using μg total DNA to μg PEI ratios of 1:1, 1:2, 1:3 and 1:6. The 1:2 and 1:3 total DNA:PEI μg ratios provided...

  6. 27 Hot Plasmids from 2016

    Type
    Blog Post
    ...expression vectors with various promoters suitable for in vivo expression, and/or produce vectors to make...of recombinant DNA technology has allowed molecular biologists to fuse fragments of DNA together in a ...prototrophic strains. The vectors were based on the original EasyClone vectors (Jensen et al 2013) allow...CDC and cloned into a third generation lentiviral transfer vector. Each gene has been FLAG-tagged for convenient...donor vectors containing mAID and either Neo or Hygro resistance markers with CRISPR/Cas vectors targeting...the authors packaged it into lentivirus and used the resulting lentiviral library to generate a population...simultaneously incorporate multiple single-strand DNA (ssDNA) oligonucleotides (oligos), and thereby rapidly...

  7. Lentivirus ddPCR Titration

    Type
    Protocol
    ...aspirate supernatant. Extract genomic DNA according to the GeneJet Genomic DNA Purification Kit instructions...., the lentiviral particles are serially diluted and used to transduce HEK293T cells. Genomic DNA is extracted... of Lentivirus Vectors You may also like... Lentivirus Colony Formation Titration Assay Lentivirus Fluorescence... (ddPCR) for lentivirus vector titration. Protocols... Protocols ddPCR Titration of Lentivirus Vectors ddPCR Titration... use droplet digital PCR (ddPCR) to titer lentivirus vectors. This protocol specifically uses primers ...Thermo Scientific, 10199-452 Reagents GeneJet Genomic DNA Purification Kit, Thermo Fisher, K0721 6-well tissue...

  8. New Optimized Genome-wide CRISPRko, CRISPRi, and CRISPRa Libraries

    Type
    Blog Post
    ...derivatives. Unmodified Cas9 causes double stranded DNA breaks that commonly result in a complete loss of...Brunello CRISPRko sgRNA library as pooled library or lentiviral prep! Dolcetto: human CRISPRi sgRNA library While...These mutations result in an inactive RNA-guided DNA binding protein (dCas9). dCas9 is then tethered to...the Dolcetto library! Now also available as a lentiviral prep. Calabrese: human CRISPRa sgRNA library ...tracrRNA was combined with the sgRNA expression vector allowing the screen to be conducted with only two...function. Find the Calabrese pooled library and lentiviral prep here! Applications for the Root and Doench...viability genes as they are subject to an increase in dsDNA breaks that can cause cell death. Thus, CRISPR inhibition...

  9. Empty Backbones - Choosing Your Perfect Plasmid Backbone

    Type
    Collection
    ...compatible Y2H vector - BD pRPR1_gRNA_handle_RPR1t - gRNA expression empty vector pFA6a vectors - Epitope ...Tet-inducible HA-tagged lentiviral vector for ORF expression pDest-565 - Gateway destination vector for bacterial... - Tet-inducible lentiviral Gateway destination vector for gene expression Lentiviral backbones made by... Tet-inducible lentiviral shRNA expression pLKO.1 - TRC cloning vector - Lentiviral shRNA expression...Blasticidin Mammalian pLX304 - 3rd gen lentiviral vector lentiSAMv2 - Lentiviral sgRNA cloning backbone pLKO.1-...Flag tag; 2NLS; pCDNA3 vector backbone, mammalian expression pENTR4-myc-nuc - pEntry vector that adds a C-terminal...pFastBac Dual LIC - Vectors from the Gradia lab pFB-LIC-Bse - Insect expression vector from Gileadi lab ...

  10. AAVs in Retinal Gene Therapy

    Type
    Blog Post
    .... At the time, adenoviral vectors were the most commonly used viral vectors for gene delivery, and they... better vector for ocular gene therapy: Adeno-Associated Virus (AAV) While other viral vectors such as...gene can be targeted to RPE cells using a lentiviral vector and a clinical trial is ongoing. Moving forward...AdV) and lentivirus (LV) are known to transduce retinal cells, AAV quickly became the vector of choice...technician in a lab using adeno-associated viral vectors (AAVs) to treat an inherited retinal degenerative...tolerate the presence of antigens (such as viral vectors) without triggering a destructive inflammatory ...retina but only transiently. However new viral vectors capable of providing long-term gene expression ...

  11. Tips for a 1st Time CRISPR User (by a 1st Time CRISPR User)

    Type
    Blog Post
    ...expressing Cas9 via transduction with Cas9 lentiviral vector, lentiCas9-Blast (52962-LV, Figure 1). Some...primers flanking my edits in genomic DNA and doing PCR on genomic DNA. The surveyor assay itself was relatively...that Surveyor nuclease cleaves double stranded DNA (dsDNA) at basepair mismatches. To use this tool to ... denature and renature the dsDNA. Upon renaturation, you will generate DNA duplexes with one strand from...performing and analyzing a gene edit using Addgene’s lentiviral CRISPR tools. Schematic overview of CRISPR...gRNA-transduced Cas9-A549 cells are used as a control. DNA was analyzed by PAGE. The basis of the Surveyor...these mismatches are indicated by the presence of DNA cleavage products. The appearance of the 428 bp band...

  12. Luciferase Plasmid Collection

    Type
    Collection
    ...cells Peter Cockerill 212936 pGL3 Basic Vector Firefly Vector for investigating regions controlling transcription... Scott Lowe 21375 pHIV-Luciferase Firefly Lentiviral vector for insertion of your gene of interest upstream... regions. Lentiviral Nadav Ahituv 212935 pGL4.84(hRlucCP/Puro) RapidResponse™ Renilla Vector for investigating...Janes 99283 pCW57.1-Luciferase Firefly TRE Lentiviral vector with dox- or tet-inducible luciferase expression...Firefly CMV Lentiviral expression of firefly luciferase Eric Campeau 18964 Luciferase-pcDNA3 Firefly CMV... Firefly EF1α Lentiviral expression of firefly luciferase Javier Alcudia 113442 pcDNA3.1 NL NanoLuc® CMV...N-terminal Nanoluc fusions using Gateway cloning. Lentiviral. Mikko Taipale 12563 PCD FL0X Firefly Insertion...

  13. Viral Vector Guides and Plasmids

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    Collection
    ...Guides and Plasmids Lentiviral Vector Guide | Lentiviral Plasmids Lentiviral vectors are versatile tools...Guides Viral Vectors Viral Vector Guides and Plasmids Selecting a Viral Vector Type Introduction to Viral...Viral Vectors Resources Viral vectors are popular tools for delivering genetic information to cells. These...the host genome. AAV Vector Guide | AAV Plasmids Adeno-associated viral (AAV) vectors are commonly used ... Gamma-retroviral Vector Guide | Gamma-retroviral Plasmids Gamma-retroviral vectors, often referred to...Retroviral vectors are popular for making stable cell lines or clinical use. Adenoviral Vector Guide | Adenoviral...applications as vaccines. Selecting a Viral Vector Type Each viral vector type exhibits different properties and...

  14. Export Control Regulations Lifted for Plasmids Containing Rabies and VSV-G Genes

    Type
    Blog Post
    ...producing lentiviral and MuLV retroviral particles. Use in conjunction with a packaging vector such as ...as pCMV-dR8.2 dvpr (lentiviral) or pUMVC (MuLV retroviral). Bob Weinberg pMD2.G VSV-G envelope expressing...paper for more information Miguel Sena-Esteves pcDNA-SADB19L Contains rabies viral RNA polymerase; ...

  15. Four Ways to Package Transgenes That Exceed the Size Limit of Adeno-associated Virus

    Type
    Blog Post
    ...roughly half the packaging limits of lentiviral and adenoviral vectors. While many transgene will fit within... overviews of three split vector strategies plus one additional split vector strategy for increasing the...homology directed repairs (HDR). Split AAV vector approaches Split vector approaches increase the size of the...copies of both vectors. Examples of use: Halbert et al., 2002 used an overlapping vector with 440 or 1000bp... hybrid vector, as measured by β-galactosidase activity, was 81% of that from a single vector, but significantly...ideal split vector system. Overall the efficiency of transgene expression from split vectors has been inconsistent..., Duan D (2008) A Hybrid Vector System Expands Adeno-associated Viral Vector Packaging Capacity in a Transgene-independent...

  16. Hot Plasmids and Viral Preps - September 2021

    Type
    Blog Post
    ...known or suspected to be involved in DNA damage response and DNA repair.  A new CRISPR pooled library ...only required three components: a master mix, the DNA of interest, and cell extract.    Graphical...They allow single-nucleotide modifications to edit DNA without requiring a double-stranded break. The Xiao... a) Cells are transfected with three different vectors and flow cytometry is used to sort populations ... 10 guide RNAs per gene. The lab modified the lentiviral backbone to encode the CD90.1 selection marker...

  17. Plan Your Experiment

    Type
    Collection
    ...viral vectors and their production, see our viral vector guides . Lentiviral vectors Lentiviral vectors... in-stock lentiviral preps . AAV vectors AAV vectors offer some advantages for viral vector delivery. ...delivery, lentiviral and adeno-associated virus (AAV) vectors are the most common. Using viral vectors takes... CRISPR plasmids . Delivery Using Viral Vectors Viral vectors are a great option, as they offer high expression...CRISPR components) integrates into the genome. Lentiviral vectors are a good option for difficult-to-transfect...don’t discuss CRISPR screens in this guide, lentiviral vectors are a common choice for conducting genome-wide.... Both lentiviral and AAV vectors are safe to use in the lab, but AAVs exhibits the lowest immunogenicity...

  18. Meet the New Addgene Board Members!

    Type
    Blog Post
    ...Baradaran’s expertise in viral safety, gene therapy vector development, vaccine development, and assay technologies...she says. In 11th grade she heard a talk about Z DNA that inspired her to major in molecular biology in...Harvard Medical School, he developed the first lentiviral microRNA-based system for polymerase II-regulated...Melina Fan and former Associate Director of Viral Vectors Karen Guerin for the journal Molecular Therapy....

  19. Choosing Your Perfect Empty Backbone

    Type
    Blog Post
    ...protein. Mammalian vectors One of our most popular mammalian expression vectors, c-Flag pcDNA3, comes from the...will need accessory packaging vectors to generate the virus. Bacterial vectors This system is great for overexpressing...cases bacterial expression vectors are inducible (e.g. pBAD LIC cloning vector (8A)) and have an epitope...folding assays. Yeast vectors There are several types of yeast expression vectors. Consider your applications...shuttle vectors. You may prefer the more traditional approach of an autonomously replicating vector that ...conduct your custom experiment. Fly vectors The vast majority of fly vectors are based on p-element, random... to reporter vectors (e.g. pBV-Luc) and gene silencing (e.g. pLKO.1 - TRC cloning vector). Check out our...

  20. Tetracycline Inducible Expression

    Type
    Collection
    ...plasmid. A single vector approach may not have as much flexibility and control as a dual vector design, but ...popular empty backbones, expression vectors for transactivators, and vectors for other applications, or search...Ptet, miniCMV Howard Gu 44012 pInducer20 Lentiviral Tet-On vector for inducible expression. See article ...Xiaojun Lian 171123 pLVX-TetOne-Puro-GFP Lentiviral Tet-On vector for inducible expression of EGFP Tet-On...TRE3GS Jason Sheltzer 11651 pLVUT-tTR-KRAB Lentiviral Tet-On vector for inducible expression of transgene ...Aebischer , Didier Trono 41393 pCW57.1 Lentiviral Tet-On vector for inducible expression; insert with ...Tight TRE David Root 100521 pCW57.1-MAT2A Lentiviral Tet-Off vector for repressible expression with blasticidin...
Showing: 41 - 60 of 111 results