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We narrowed to 294 results for: expression vector

Showing: 121 - 140 of 294 results

  1. Hot Plasmids - August 2020

    Type
    Blog Post
    Published
    Aug. 18, 2020, 1:15 p.m.
    ...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...

  2. 5 Tips for Troubleshooting Viral Transductions

    Type
    Blog Post
    Published
    Aug. 11, 2016, 2:23 p.m.
    .... 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...

  3. Tips for Screening with Yeast Two Hybrid Systems

    Type
    Blog Post
    Published
    Oct. 22, 2015, 2:30 p.m.
    ...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...

  4. DAP arrays for multiplex gene editing

    Type
    Blog Post
    Published
    July 7, 2022, 1:15 p.m.
    ...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...

  5. Hot Plasmids and Viral Preps - March 2021

    Type
    Blog Post
    Published
    March 23, 2021, 1:15 p.m.
    ...-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...

  6. CRISPR/Cas9 FAQs Answered!

    Type
    Blog Post
    Published
    March 13, 2014, 4:08 p.m.
    ...use double nickase, you could express two spacers and use PX335 to express the Cas9n (nickase). The concept...start site to have high expression, so adding this G could help with expression, though it is possible ...you need to have four spacers co-expressed and using PX335 to express the Cas9n (nickase). Tips for using...concept of the double nickase system is that you can express two different chimeric gRNAs with the Cas9 nickase...oligo when cloning my target spacers into the PX330 vector backbone? A2: There is no need to add the NGG PAM...backbone that uses the human U6 promoter to drive expression, is it necessary to add a G nucleotide to the...possible for the plasmid to still express without the G. Because the G is only one base, the Zhang lab usually...

  7. Hot Plasmids: Spring 2025

    Type
    Blog Post
    Published
    June 3, 2025, 1:15 p.m.
    ...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...

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

    Type
    Blog Post
    Published
    June 11, 2020, 1:34 p.m.
    ...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...

  9. PiggyBac-ing Through the Genome Editing Field

    Type
    Blog Post
    Published
    May 31, 2016, 3:30 p.m.
    ... 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...

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

    Type
    Blog Post
    Published
    July 18, 2017, 12:41 p.m.
    ...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...

  11. CRISPR 101: Epigenetics and Editing the Epigenome

    Type
    Blog Post
    Published
    June 24, 2020, 5:45 p.m.
    ...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...

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

    Type
    Blog Post
    Published
    Oct. 29, 2015, 2:30 p.m.
    ...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...

  13. Plasmids 101: Repressible Promoters

    Type
    Blog Post
    Published
    Dec. 20, 2022, 2:15 p.m.
    ...to be expressed. These permit exquisite control of gene expression and tracing of gene expression across... basal expression. Co-expression of repressor QS along with QF leads to intermediate expression from QUAS...transgene permits directed expression of that gene in cells that also express GAL4. Usage of this system...UAS to examine the expression of reporter genes, or to combine reporter expression from one promoter with...basal state, and co-expression of the inducer QF can increase QUAS driven expression by ~3,300-fold in ... Shen. “Controlling gene expression with the Q repressible binary expression system in Caenorhabditis ...gene, it therefore determines the timing of gene expression and largely defines the amount of recombinant...

  14. Hot Plasmids February 2024

    Type
    Blog Post
    Published
    Feb. 20, 2024, 2:45 p.m.
    ...escape. Combining Cas9-PAGE with lentiviral sgRNA expression yielded editing of 80–90% in human cell lines...structured illumination microscopy in HeLa cells expressing COX8a-mStayGold. Scale bars 10 μm (main), 1 μm...developed an engineered adeno-associated virus (AAV) vector named AAV.CAP-Mac for non-invasive systemic gene...bias in mature marmosets. CAP-Mac also robustly expressed fluorescent reporters in green monkeys and cultured...cultured human neurons, with broader neuronal expression than AAV9. This new capsid will be a valuable ...N.C., Goeden, N. et al. Adeno-associated viral vectors for functional intravenous gene transfer throughout...region VIII of AAV9-X1.1 to that of AAV1. This viral vector has brain endothelial cell-specific tropism, with...

  15. Three Tips for Preventing Viral Plasmid Recombination in Your Samples

    Type
    Blog Post
    Published
    Sept. 14, 2021, 1:15 p.m.
    ...plasmids are formed. Viral vectors are prone to recombination In lentiviral vectors, recombination often occurs...Improved recombinational stability of lentiviral expression vectors using reduced-genomeEscherichia coli. BioTechniques...problems that I help troubleshoot involves viral vectors and plasmids with repeating sequences. Repeating.... Over time, bacteria containing the recombined vector backbone will also outgrow bacteria that contain...into two smaller plasmids. The ~3.7 kb recombined vector backbone contains the antibiotic resistance gene...samples? Plasmid recombination can affect viral vectors (LTRs and ITRs), plasmids with multiple polyA sites...recommendations from the depositing lab. If your vector is large (>10 kb) or has a low copy number, try...

  16. How to Deposit Your Plasmids with Addgene

    Type
    Blog Post
    Published
    Feb. 27, 2024, 2:15 p.m.
    ...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...

  17. FastCloning: A Newer, Simpler Cloning Technique

    Type
    Blog Post
    Published
    Nov. 1, 2016, 2:30 p.m.
    ... insert and vector are first PCR amplified. The forward primer used to amplify the vector is located in...your vector and insert. Once you’ve confirmed that the PCR was successful, mix together the vector and ... PCR with vector specific primers to confirm that the products are the correct size (vector + insert)....specific sequences for base pair overhangs, insert and vector purification, and, most importantly, excessive ..., it does not require purification of insert or vector products, nor does it require the use of specific...process in brief In FastCloning, you PCR both the vector and the insert in separate reactions and then combine...bases that are complementary to the overlapping vector ends. See an overview of the PCR process and the...

  18. Hot Plasmids September 2018 - Optogenetics, RNA Localization, Fluorescent Protein, and Base-editing Tools

    Type
    Blog Post
    Published
    Sept. 18, 2018, 1:33 p.m.
    ...available in a mammalian expression vector (pcDNA3) and a bacterial expression vector (pRSETB). Fabritius...APOBEC. To achieve this they created a lentiviral vector system that monitors both APOBEC- and Cas9-mediated...ACE) in real-time. The reporter constitutively expresses eGFP to indicate reporter abundance, and mCherry...both halves of the modified EB1 protein are overexpressed in cells, microtubule growth can be switched...

  19. PITChing MMEJ as an Alternative Route for Gene Editing

    Type
    Blog Post
    Published
    Feb. 23, 2016, 3:30 p.m.
    ...cloned into a vector containing Cas9 and the PITCh-gRNA. Step 3: Contransfect the PITCh vector with the vector...repair, making it easier to construct targeting vectors. Addgene depositor Takashi Yamamoto’s lab has harnessed...just upstream of a stop codon. Briefly, the PITCh vector should be constructed with 5’ and 3’ microhomology...PITCh Step 1: Generate microhomologies in the PITCh vector ~20 bp 5’ and 3’ microhomologies are added to ..., and this construct is inserted into the PITCh vector via In-Fusion or SLIC cloning. Step 2: Design an... vector carrying Cas9 and the PITCh- and locus-specific gRNAs Step 4: Select for puromycin resistant cells...plasmids available from Addgene are perfect for expressing GFP from a given promoter, and the technique ...

  20. What the HEK?

    Type
    Blog Post
    Published
    April 18, 2023, 1:15 p.m.
    ...derivatives of HEK-293, the most commonly used line expresses SV40 large T antigen and is thus dubbed HEK-293T...packaging cells’ for viral production because they express the SV40 large T-antigen. This allows them to replicate...receptors and ion channels are often exogenously expressed and studied in HEKs due to the cell line’s ability...produce these proteins and its relatively low expression of native channels.  No matter what your research...factors that could skew research results (gene expression, growth rates, etc.). Thus, it’s important to...Platform to Produce Recombinant Proteins and Viral Vectors. Front Bioeng Biotechnol. (2021) (9):796991. DOI...10 Tips for Mammalian Cell Culture Adenoviral Vector Production and Troubleshooting Additional resources...
Showing: 121 - 140 of 294 results