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We narrowed to 407 results for: expression vector

Showing: 121 - 140 of 407 results

  1. Protocol - Bacterial Transformation

    Type
    Protocol
    ...plasmids (even those designed for mammalian cell expression) carry both a bacterial origin of replication...approximately 10-fold lower for ligation of inserts to vectors than for an intact control plasmid. Incubate the...

  2. The 10 Most Distributed Plasmid Technologies in Addgene's First 10 Years

    Type
    Blog Post
    ...backbones for shRNA expression, such as pLKO.1 puro and pLKO.1 – TRC cloning vector. Lentiviral Plasmids...requests grow for lentiviral packaging and expression vectors, including pMD2.G and psPAX2. Actually, these... Our first popular "kit," Andrew Fire's set of vectors for C. elegans research was our most requested ...empty backbones such as pBABE-puro for retroviral expression and pLVTHM, a lentiviral plasmid for mammalian...

  3. Viral Vectors 101: Producing Your rAAV

    Type
    Blog Post
    ...Transgene Expression in Cultured Cells Using Unpurified Recombinant Adeno-Associated Viral Vectors [Preprint....  In this helper-free system, HEK293T cells, expressing Ad E1A and E1B, can be transfected with your ...you’ll need to determine the titer of your viral vector solution. A variety of titering methods have been...about common AAV applications by reading our Viral Vectors 101 series!  This post was originally written by...rapid and cost-effective manipulation of gene expression in the CNS. Molecular Neurodegeneration, 15(1...the Stability and Inactivation of AAV Serotype 1 Vectors. Human Gene Therapy Methods, 28(1), Article 1. ...(2021). Inactivation of Adeno-Associated Viral Vectors by Oxidant-Based Disinfectants. Human Gene Therapy...

  4. Viral Vectors 101: AAV Serotypes and Tissue Tropism

    Type
    Blog Post
    ...Viral Vectors 101: An Introduction to AAV Viral Vectors 101: AAV Variables that Matter Viral Vectors 101...choice of AAV vector for your experimental system. AAV1 AAV1 was the first viral vector approved for gene...A viral vector that can target specific tissues, even when administered systemically, without causing...causing disease? Recombinant adeno-associated viral vectors, or rAAVs, can sound almost too good to be true...components. Created with BioRender.com. AAV vectors have both distinctive capsids and genomic structures...specific cells. This protein shell packages the vector’s genetic material, and variations between serotypes...target species or even strains. Pro tip! Getting a vector into a particular cell type depends largely on ...

  5. 15 Hot Plasmids from 2017

    Type
    Blog Post
    ...low off-target activity; mammalian expression vector Rees H, et al. Nature Communications. 2017. PubMed...and ribozyme systems for multiplex gRNA expression. Expression of toolkit components can be controlled...gene editing. The bacterial and mammalian expression vectors for these CRISPR inhibitors are listed in...Species Bacterial Expression Construct SpCas9 Inhibition (E. coli) Mammalian Expression Construct SpCas9...TagRFPt, EGFP, mVenus, mCerulean3, mKOFP2) and empty vectors that have standard cloning sites or gateway compatible...neuronal tools to the expanding gateway compatible vectors currently shared amongst the zebrafish community... (induced pluripotent stem cells) through the expression of transcription factors OCT3/4, SOX2, KLF4, ...

  6. Tips for arabidopsis transformation

    Type
    Blog Post
    ...Addgene  Choosing an expression system Choosing an appropriate expression vector is the first step in .... (2007) ‘Improved Gateway Binary Vectors: High-Performance Vectors for Creation of Fusion Constructs ...from the Tsuyoshi Nakagawa lab are binary gateway vectors and can be used for promoter swapping and generating...for visual selection of transformants based on expression of RFP in the seed (Emami, Yee and Dinneny, 2013...many insertions can cause artifacts from over-expression and it is likely that such a construct will be... ‘A robust family of Golden Gate Agrobacterium vectors for plant synthetic biology’, Frontiers in Plant...sweet potato contains Agrobacterium T-DNAs with expressed genes: An example of a naturally transgenic food...

  7. Plasmids 101: Terminators and PolyA signals

    Type
    Blog Post
    ... terminator will suffice. Many commercial expression vectors use double terminators to reduce unwanted...Regulatory Element Enhances Expression of Transgenes  Delivered by Retroviral Vectors. Journal of Virology. ...Plasmids designed to express genes in a given host cell type are generally broken down into two broad...on promoter strength as a determinant of gene expression levels, the terminator also plays an important...variability in RNA half-life, and ultimately gene-expression. Polyadenylation is the post-transcriptional ...terminators are not usually employed in plasmid-based expression systems, so these will not be detailed here, ...provided at the end.  Nearly all common bacterial expression plasmids use Rho-independent terminators, which...

  8. Technique: Probe Phage Genomes for Host Binding Proteins

    Type
    Blog Post
    ...point for this experiment. Find Bacterial Expression Vectors in the FX Cloning System Kit  Step 3: Probe...Clone Your Gene of Interest Into a Variety of Expression Vectors Deposit Your Microbiology Plasmids ...ligate into a bluntly digested vector designed for protein overexpression in E. coli, such as pET30a. If...homology-based searches return no candidates. Our strategy: Express random fragments of the phage genome and probe ...primers bordering the multiple cloning site of the vector. In order to reduce the chances of some clones ...plate) across 20 fresh selective plates. Step 2: Express random phage DNA fragments in E. coli Now that ...and exposed to IPTG in order to induce protein expression. Cells on the membranes will then be exposed ...

  9. 5 Tips for Troubleshooting Viral Transductions

    Type
    Blog Post
    .... Viral vectors undergo DNA rearrangements. Depending on the composition of the viral vector and the ... or resistance marker expression following transfection, higher-level expression after approximately 72...about Lentiviral Vectors Additional Resources on the Addgene.org Check out Our Viral Vector Guide Pages ... 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...

  10. Hot Plasmids - August 2020

    Type
    Blog Post
    ...easier to clone, express, and purify compared to antibodies. Nanobodies are easily expressed in the cytosol...#140702) The pMAK461-64 plasmids were expressed in SHuffle T7 Express competent E.coli (NEB) using IPTG induction...approaches rely on continual expression of both dCas9 and the gRNA. This expression can vary significantly ...fluorescently labeled piggyBac vectors for uniform and sustained expression of single or multiplexed gRNAs...and track cells expressing both dCas9 and gRNAs. The multi-gRNA piggyBac vector integrates into ... the mCitrine gene. mCherry is constitutively expressed. Cells are then cultured and FACS sorted. Image...Histone 2A or 2B can result in bright targeted expression while minimizing non-specific labeling. For labeling...

  11. Tips for Screening with Yeast Two Hybrid Systems

    Type
    Blog Post
    ...Yeast Expression Vectors Find Bacterial Two Hybrid Plasmids Read our Blog Post About Yeast Vectors Read...activation. Vary the expression levels of bait and prey proteins - overexpression can lead to false interactions...During this period, proteins were successfully expressed as fusions that retain their individual activities...see Figure 2a) (4). These fusions are then co-expressed in a yeast strain that also contains a reporter...bait”, Figure 2b) cannot activate reporter gene expression on its own, but, if the two POI’s interact, this...the promoter of the reporter gene to activate expression (Figure 2c). The reporter gene then allows detection...strains are used to select for binding-dependent expression of the HIS3 gene, which is required for histidine...

  12. DAP arrays for multiplex gene editing

    Type
    Blog Post
    ...promoter, which is used to drive the expression of the array. After expression, the individual gRNAs are released...number of gRNAs that can be packaged into viral vectors for potential in vivo applications or multiplex...demonstrated that the tRNA-gRNA array can drive the expression of itself, without using any additional promoter...unprecedented number of genes with a single viral vector.   Fig. 1: Schematic of multiplex gene ..., that rely on an upstream lengthy promoter to express the gRNA array, DAP arrays use tRNA itself, as ...is scalable and efficient in plasmid and viral vectors. In applications when repetitive tRNA sequences...resources on the Addgene blog CRISPR 101: Multiplex expression of gRNAs Four Base Editing Reporters to Monitor...

  13. Hot Plasmids and Viral Preps - March 2021

    Type
    Blog Post
    ...-baToN, “sender” cells express GFP on the surface and “receiver” cells express an anti-GFP nanobody on...response 1 (TIR1) expressed under a desired promoter with a specific spatiotemporal expression pattern and ... nanobody cross-reacts with BFP, sender cells expressing surface BFP generated BFP-labeled receiver cells...visualized.  The tool-kit consists of highly useful vectors compatible with Gibson and Saptrap cloning systems...for delivering CRISPRi components by a single AAV vector are now available from Tonia Rex's lab.  vfCRISPR...Jacobs-Wagner’s lab produce different levels of dCas9 expression and contain different antibiotic resistance markers...from Hongkui Zeng's lab are available for dual expression of cytoplasmic-tdTomato and Synaptophysin-GFP...

  14. Chemogenetics vs. Optogenetics: Which Method Should I Choose?

    Type
    Blog Post
    ...more about optogenetics New to viral vectors? Browse our Viral Vectors 101 blog posts Learn more about precise...Addgene.org Browse all viral vectors available at Addgene Visit our viral vectors guide Find our chemogenetics...optogenetic and chemogenetic tools can have their expression targeted to specific tissues, cell types or even...restricted to illuminated region Impacts all cells expressing receptors Invasiveness Mice require permanent...

  15. Hot Plasmids: Spring 2025

    Type
    Blog Post
    ...backbones special: Strong expression: A CMV promoter drives robust gene expression in mammalian cells. Flexible... plasmid set for expressing tagged proteins, and stay tuned for mammalian expression and AAV plasmid sets...therapies. Using AAV vectors with enhancers that drive cell-type-specific expression has been a promising...) Fluorescence from axonal boutons of neurons expressing iGluSnFR3, iGluSnFR4f, and iGluSnFR4s. C) Spike-triggered...fusions for membrane display, they recommend the NGR vectors for most experiments. These new tools promise to... toolkit is the empty plasmid backbone: the DNA vector that contains everything needed to replicate inside...share our very own set of empty, HA-, and EGFP-expressing lentiviral backbones designed right here at Addgene...

  16. PiggyBac-ing Through the Genome Editing Field

    Type
    Blog Post
    ... minimize expression levels of your gene of interest, as needed. Controlling the expression levels of ...quickly transfer a DNA sequence from the transposon vector to one of many TTAA sequences distributed throughout...encourage you to deposit your piggyBac™ transposon vectors with us to help us expand this useful resource....may be particularly useful when the gene being expressed has threshold effects, such as with signaling ... can also mitigate the deleterious effects of expression of certain genes, such as those that have negative...edit of interest. With the larger piggyBac™ donor vector, it is possible to make the ITR and selection cassette...been shown to cause genomic abnormalities when expressed. This lack of off-target effects is likely due...

  17. How to Deposit Your Plasmids with Addgene

    Type
    Blog Post
    ...the vector backbone you cloned your insert into and fill out the appropriate boxes with the vector backbone...backbone manufacturer, vector backbone size, total size of your vector + insert, any modifications you ...insert in E. coli, then this is a Bacterial Expression vector. If your plasmid can be classified in multiple...lentiviral vector to be used in mammalian cells, check Lentiviral and Mammalian Expression). This information... location, while shRNAs are used to knockdown expression of mRNAs with complementarity to the shRNA sequence...provide any full or partial plasmid sequences and any vector maps. The more sequence data available, the better... was a part of the insert (when cloned into the vector backbone). If your plasmid has more than one insert...

  18. Addgene-Seeding Labs Plasmid Grant: Accelerating Science Globally - 5 Scientists, 4 Countries & 3 Continents!

    Type
    Blog Post
    ...the light‐inducible bacterial expression system pDawn to clone and express newly identified proteins and...project the expression of isolated bacterial proteins can be achieved using a bacterial expression system ...dietary supplements! Development of a bacterial expression system for renewable biotechnology products Dr...focuses on molecular biotechnology for improving expression of nutritionally and therapeutically significant...cannot produce them. By using yeast integration vectors (see for example this yeast cloning kit) she intends...system such as the pET GFP LIC cloning vector and the pOKD plasmids. The recombinant Cry proteins will then...H2O2 probe is pQE-30-HyPer3 (2) which he will co-express together with a plasmid encoding glycine oxidase...

  19. CRISPR 101: Epigenetics and Editing the Epigenome

    Type
    Blog Post
    ...the lab used this vector in chick embryos, it also functions in mammalian expression systems.   Christopher... are an additional layer of control over gene expression that go beyond genomic sequence. Dysregulation...modifications are correlated with alterations in gene expression, but causality and mechanisms remain unclear....that help package DNA, were associated with gene expression or silencing. To alter DNA modifications, researchers...Charles Gersbach’s lab has deposited mammalian expression constructs including pcDNA-dCas9-p300 Core and...well as pLV-dCas9-p300-P2A-PuroR for lentiviral expression. Figure 1: dCas9-p300 adds H3K27ac marks...methylation and activate transcription. A lentiviral vector with the same modifier, Fuw-dCas9-Tet1CD, is available...

  20. Plasmids 101: A Brief History of Plasmids and an Improved eBook!

    Type
    Blog Post
    ...cloning vectors such as pBR322, pACYC, and pUC were developed to provide higher copy number vectors that ...added to the "Plasmids that Glow" and "Viral Expression Vectors" chapters, and more pictures and pro tips...successful, pSC101 became the first plasmid cloning vector and molecular biology was never the same. Over ...replication and topology, and cloning and gene expression. Just released: Addgene's Plasmids 101 (2nd...
Showing: 121 - 140 of 407 results