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We narrowed to 370 results for: Cre expression vector

Showing: 121 - 160 of 370 results

  1. Viral Vectors 101: Optogenetic Tools

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    Blog Post
    .../cells expressing the optogenetic tool causing a change in cellular activity or gene expression. Experimental...control everything from electrical activity to gene expression, and even CRISPR based gene editing. But, how...hyperpolarize the cell (Mattis et al., 2012). Bicistronic vectors combine excitation and inhibition by using both...confounding issues associated with heterologous expression of engineered receptors. OptoXRs are great for...candidates for your experiments.  Wavelengths Most vectors have an optimal wavelength to use but can be activated...Experimental output is measured and quantified. Created with BioRender.com.   Optogenetic tools Types...passing ions against the concentration gradient. Created with BioRender.com.   Opto-G-protein coupled...

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

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    ...requests grow for lentiviral packaging and expression vectors, including pMD2.G and psPAX2. Actually, these... In 2006, we saw an increase in popularity of empty backbones for shRNA expression, such as pLKO.1 puro... Our first popular "kit," Andrew Fire's set of vectors for C. elegans research was our most requested ...puro and pLKO.1 – TRC cloning vector. Lentiviral Plasmids -  If you're familiar with the pLKO plasmids ...empty backbones such as pBABE-puro for retroviral expression and pLVTHM, a lentiviral plasmid for mammalian...was Addgene's first distributed plasmid and was created in the lab of my thesis advisor, Dr. Bruce Spiegelman...requested. Ubiquitin - One particular gene we saw increased interest in from 2008 to 2009 was ubiquitin. The...

  3. Viral Genome Sequencing - A One Stop Shop for AAV Identity and Serotype Confirmation

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    Blog Post
    ...the percent of recombination present in Cre-dependent vectors and a script that can confirm the serotype... while we think of the AAV vector genomes as containing just the expression cassette that we want, it ... our viral vectors related blog posts Read our blog post on AAV titers Browse viral vectors protocols ...accommodate adeno-associated viral vector (AAV) samples and created a simplified process, termed viral...methods for AAV quality control Historically, AAV vector QC has been sparse including only titration of ... of quality control is sufficient but for viral vector core facilities producing a large number of viral...demonstrated that NGS is an effective tool to confirm AAV vector genome identity. These studies used SMRT sequencing...

  4. Tips for arabidopsis transformation

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    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 in Transgenic...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...

  5. Viral Vectors 101: AAV Serotypes and Tissue Tropism

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    Blog Post
    ...Viral Vectors 101: An Introduction to AAV Viral Vectors 101: AAV Variables that Matter Viral Vectors 101...: Diagram of AAV components. Created with BioRender.com. AAV vectors have both distinctive capsids...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...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 ...viral capsid, but specificity doesn’t end there: expression from rAAVs also depends on your choice of promoter...

  6. Plasmids 101: Terminators and PolyA signals

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    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. ...polyadenlyation. Mammalian expression plasmids are primarily used to create mRNA and the commonly used...Internal Polyadenylation Signal Substantially Increases Expression Levels of Lentivirus-Delivered Transgenes...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 ...

  7. Viral Vectors 101: Producing Your rAAV

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    Blog Post
    ...Transgene Expression in Cultured Cells Using Unpurified Recombinant Adeno-Associated Viral Vectors [Preprint...minimally purified secreted rAAV for rapid and cost-effective manipulation of gene expression in the CNS. Molecular....  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...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...

  8. Technique: Probe Phage Genomes for Host Binding Proteins

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    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 ...T4 DNA polymerase (to create blunt ends) and ligate into a bluntly digested vector designed for protein...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

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    .... 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. DAP arrays for multiplex gene editing

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    Blog Post
    ...promoter, which is used to drive the expression of the array. After expression, the individual gRNAs are released...packaged into viral vectors for potential in vivo applications or multiplex CRISPR screening.   Using tRNAs...CRISPR screenings that would perturb the unprecedented number of genes with a single viral vector.   ...demonstrated that the tRNA-gRNA array can drive the expression of itself, without using any additional promoter..., 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...

  11. Hot Plasmids and Viral Preps - March 2021

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    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...

  12. Hot Plasmids: Spring 2025

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    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...

  13. CRISPR/Cas9 FAQs Answered!

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    Blog Post
    ...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...

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

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    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...Scientists continue to engineer opsins to operate at increasingly faster kinetics. The opsin ChETA, for example...regions of a neuron. Optogenetic approaches excel at discrete cell or tissue-subregion targeting down to the...

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

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    ...project the expression of isolated bacterial proteins can be achieved using a bacterial expression system ...fast endophyte screening system and plans to employ the light‐inducible bacterial expression system pDawn...LIC cloning vector and the pOKD plasmids. The recombinant Cry proteins will then be screened for toxicity...dietary supplements! Development of a bacterial expression system for renewable biotechnology products Dr...pDawn to clone and express newly identified proteins and enzymes. These compounds will later get tested for...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...

  16. CRISPR 101: Epigenetics and Editing the Epigenome

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    ...the lab used this vector in chick embryos, it also functions in mammalian expression systems.   Christopher...pcDNA3.1-MS2-Tet1-CD to decrease methylation and activate transcription. A lentiviral vector with the same modifier...Arlotto ME, et al (2018) Creation of versatile cloning platforms for transgene expression and dCas9-based epigenome... 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...

  17. Hot Plasmids - October 2020

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    ...from other regions of the cell. By combining vectors expressing different fluorescent proteins targeting ...Multiplexed Editing and screening Applications (CHyMErA) Knockout Libraries express hybrid Cas9-Cas12a gRNAs...display (FingRs), Xue Han’s lab has developed AAV vectors to selectively visualize excitatory and inhibitory...excitatory and inhibitory synapses, respectively. Once expressed in the neuron, the FingRs bind to the endogenous...targeting inhibitory or excitatory synapses or that are Cre-inducible, researchers can explore synaptic integration.... The team engineered C. elegans cell lines to express wrmScarlet1-10 unattached to any other cellular...proliferation in acute myeloid leukemia and saw that expression of H2B-FT consistently predicted the cell cycle...

  18. Plasmids 101: Repressible Promoters

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    ...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...basal state, and co-expression of the inducer QF can increase QUAS driven expression by ~3,300-fold in ...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... Shen. “Controlling gene expression with the Q repressible binary expression system in Caenorhabditis ...promoters and create sophisticated patterns of UAS-driven gene expression. To further increase the complexity...

  19. Hot Plasmids February 2024

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    ...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...

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

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    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...Plasmids 101 eBook! In keeping with our goal of creating a one-stop reference guide for plasmids we've ...

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

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    ...available in a mammalian expression vector (pcDNA3) and a bacterial expression vector (pRSETB). Fabritius...deaminases like APOBEC. To achieve this they created a lentiviral vector system that monitors both APOBEC- and...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...cobalamin fusion to the riboswitch results in increased fluorescent signal. Prior to the development of...developing a new imaging-based bacterial colony screening platform, Oliver Griesbeck’s lab set out to engineer...the red end of the spectrum (above 600 nm) and increasing the brightness. Arne Fabritius and colleagues...

  22. How to Deposit Your Plasmids with Addgene

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    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...

  23. PITChing MMEJ as an Alternative Route for Gene Editing

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    Blog Post
    ...cloned into a vector containing Cas9 and the PITCh-gRNA. Step 3: Contransfect the PITCh vector with the vector...construct targeting vectors. Addgene depositor Takashi Yamamoto’s lab has harnessed MMEJ to create a new method...if the GFP-Puro will alter expression of the gene it follows. For increased versatility, it would be advantageous...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...

  24. FastCloning: A Newer, Simpler Cloning Technique

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    Blog Post
    ... 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...

  25. X-CHIME: Context Dependent Germline Knockout in Immune Cells

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    Blog Post
    ...transduction of Cas9-expressing (Rosa26-Cas9) hematopoietic stem cells with a lentiviral vector containing a ...utilizes custom mouse strains and lentiviral expression vectors (Table 1). The four systems Name Function... pXPR_070 Violet-excited GFP (vex) Lineage-Cre; Rosa26-Cas9 S-CHIME Knocks out pairs of genes...of genes in immune cells typically involves the creation of germline knockout mice, which is time-consuming...cells can be implanted in irradiated recipients to create mice with gene knockouts in their immune systems...2019). A CRISPR-Cas9 delivery system for in vivo screening of genes in the immune system. Nature Communications...

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

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    Blog Post
    ...because they can be expressed in bacteria, while mAbs generally require mammalian expression systems. scFvs...used to screen for new antibodies. To do this, antibody variable domain sequences are expressed as scFvs... plant, and insect cells, they are most often expressed from a plasmid in bacteria (Ahmed et al., 2012...2012). One challenge of bacterial expression is the proper formation of the disulfide bond between the VL...requires an oxidizing environment to form. When expressed in the non-oxidizing cytoplasm of bacteria, scFvs...more of these approaches are often used for scFv expression in bacteria:  targeting to the oxidizing periplasm...to allow for proper disulfide bond formation.  expression in strains of bacteria that are redox mutants...

  27. Using Phosphoserine to Study Protein Phosphorylation

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    Blog Post
    ...recognized by the Sep-tRNA. 3. An expression plasmid encoding the ORF to be expressed Finally, the researcher needs...ORF of interest is cloned into one of these expression vectors, the codon for the amino acid to be replaced...non-phosphorylated serine at the TAG codon of the same expression vector used for phosphoserine incorporation (6). ...using the same expression plasmid. This is a particularly important control when creating a phosphoserine-incorporation...bacterial host strain for protein production, and an expression plasmid encoding the open reading frame (ORF)...encoding their open reading frame of interest. Two expression systems have been verified for use by the Rinehart...TOPO E17TAG GFP and pGEX-6P-1. The pCRT7 NT TOPO vector contains a mutant pSer-containing GFP that can ...

  28. With Our New Viral Service, We're Taking CRISPR Further!

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    Blog Post
    ...easy process. The cloning capacity of lentiviral vectors (~8.5 kb of insert) and the reduced titer associated...providing lentiviral preps of select lentiviral vectors from the repository for you. We perform test infections...infections with all of our viral preps and host our expression data (western blots or fluorescent microscopy...nickase for genome editing. You can also enhance expression of your gene of interest with a variety of Cas9...therefore, must be used to infect a cell line already expressing Cas9. As John Doench (Broad Institute and member...Page Check Out Our CRISPR Guide Pages Find Viral Vectors ...CRISPR pooled gRNA libraries that can be used to screen for genes involved in a wide variety of cellular...

  29. CRISPR Kinome Libraries Available: Pooled and Individual Plasmid Formats

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    ...pools are available in two vector formats: all-in-one (lentiCRISPRv2, expressing both Cas9 and an individual...individual gRNA from this single vector) and guide-only (lentiGuide expressing only an gRNA, for use with cells... useful for imaging or gene expression assays necessitating the screening of individual plasmids separately...Library Screening Why the kinome? While properly designed perturbations – in the case of screens using ...at the Broad Institute, John Doench. A genetic screening project can be a tremendous undertaking, producing...technology, gRNAs to knockout genes – are essential screening components, the model system and the assay are...is to do it huge, there are times when a more incremental approach is prudent. What's in the CRISPR kinome...

  30. Why Add Sucrose? Improved Yields for Adeno-associated Virus Preparation

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    ...guide to viral vectors Find all of our viral vector blog posts Download our viral vectors 101 eBook Additional...pressure on the production of retroviral vectors: Enhancement in vector stability." Biotechnology and bioengineering...been actively investigating ways to improve viral vector production. Whether redesigning viral packaging...membrane When packaging adeno-associated viral vectors (AAV) not all serotypes are equal; while some serotypes... of a fluorescent dye and transduction of GFP-expressing AAV particles we did not observe any difference...we believe that this approach can be adapted by vector cores and laboratories alike as an easy and inexpensive...Efficient serotype-dependent release of functional vector into the culture medium during adeno-associated...

  31. Making CRISPR Plasmids Using Fragmid

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    Blog Post
    ...modular vector assembly process designed for a quick, plug-and-play approach to cloning CRISPR vectors (McGee...for each module, generate the vector file, and quickly clone their vector of interest. And it truly is ...destination vectors for various delivery methods, along with compatible inserts. All destination vectors contain...heavy lifting out of CRISPR vector design, making it easier to get the vectors that are just right for your...Fragmid destination vectors currently available. Table 1: Fragmid destination vectors Organism Delivery...designing a CRISPR vector the Fragmid way!    Figure 1: Selecting your vector type on the Fragmid...plasmid and generating the vector file on the Fragmid website.   If the vector is to your liking, your...

  32. Tag Your Favorite Yeast Genes with Ease

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    Blog Post
    ...codon optimized specifically for expression in yeast. These tagging vectors are based on previously described...interest. These fluorescently tagged genes are expressed under native conditions and allow scientists to...described pFA6a-link vectors and include a Kan, SpHIS5, or CaURA3 selection marker. Lee et al.(1) assessed...from these yeast-optimized fluorophore tagging vectors for your single or multi-color imaging experiments...artifacts sometimes associated with plasmid-based overexpression. Tim Formosa, at the University of Utah, has...multiple cloning site in place of the epitope tag for creation of your own unique protein fusions. This collection..., including full and partial gene deletion, overexpression (by promoter substitution), and tagging at ...

  33. Cloning Mammalian Cells with the Agarose Method

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    ...Fluorescent Protein Plasmids Find Mammalian Expression Vectors ...that overexpress genes of interest; more recently we have also been using Addgene CRISPR vectors to generate... enzyme assays to select both over- and under-expressing clones. If GFP-encoding cDNA is co-transfected...and PCR, depending on the level of endogenous expression. We highly recommend the use of an electronic...ampules each of 4 clones with different levels of expression of our gene of interest. Advantages of the agarose...isolate individual cell clones with different expression characteristics from a population of transfected...Challenges of Cell Culture Plasmids 101: Mammalian Vectors Resources on Addgene.org Check Out Our Protocols...

  34. Channelrhodopsins with improved light sensitivity for minimally-invasive optogenetics

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    Blog Post
    ... injection of an AAV expressing a ChRger. In both experiments, neurons expressing ChRgers produced stronger... what was seen with ChRger2 expressing mice, while ChR2(H134R) expressing mice did not have this self-...Addgene.org Browse all viral vectors available at Addgene Visit our viral vectors guide Find our chemogenetics...to fire when light is shined on them, but the expression of light-gated ion channels such as channelrhodopsins...When light shines on excitatory channelrhodopsin-expressing neurons, the channelrhodopsins respond by opening...sequence and structures that determine membrane expression and plasma membrane localization of transmembrane...and ChRger3, were then characterized further by expression in cultured neurons and mouse brain slices collected...

  35. What's New in CRISPR - September 2019

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    ...tools for your research! This time: GeneWeld vectors to create knock-ins CasX Drug inducible CRISPR/Cas9 ...libraries for cancer research Creating knock-ins with GeneWeld vectors GeneWeld from Jeffrey Essner’s...’s lab uses CRISPR/Cas9 and a donor vector to create knock-ins of fluorescent proteins, fusion proteins...cargo (ex: fluorescent protein gene) on the donor vector. Therefore, targeting requires two sgRNAS: one ...site and a universal sgRNA that targets the donor vector sequence. Homology arms that flank the donor plasmid... Read the paper in bioRxiv Find the GeneWeld vector kit CasX, a small Cas protein, is active in E....active and inactive CasX for E. coli and human expression. Read the paper in Nature Find the CasX plasmids...

  36. Top Requested Plasmid of 2017 - pMD2.G

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    Blog Post
    ...2016 were CRISPR plasmids), but so are lentiviral vectors, many of which are used to deliver Cas9 and other....G from Didier Trono’s lab! Lentivirus envelope vector pMD2.G This plasmid has been requested nearly 6,000... 6,000 times (over 800 orders in 2017) and expresses the VSV-G envelope protein under the control of the... our Viral Production Process Find other Viral Vectors ... This plasmid has been used in studies of the secretome, ubiquitin ligase, cellular senescence, autophagy...

  37. Hot Plasmids - June 2019 - Optogenetics, Acoustic Reporter Genes, microRNAs, and the CRISPR-Cas9 system CHIME

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    ...manner. They designed the pXPR_053 lentiviral expression vector, which contains an sgRNA insert site and a...pXPR_053. They expressed this construct in bone marrow stem cells from Cas9-expressing mice, and used ... microbe travels through a host organism, the expression of some genes can change depending on the location...reporter gene (ARG) plasmids to better monitor gene expression in vivo using ultrasound. These ARG plasmids .... 2018. PubMed PMID: 29300010. miRNA reporter vectors shed new light on mechanisms of miRNA-mediated ...important regulators of post-transcriptional gene expression across the majority of cellular pathways. Despite...target site. One fluorescent protein (EBFP2) is expressed in an unregulated fashion and thus a marker for...

  38. What's New in CRISPR - Spring 2019

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    ...doxycycline, but it can also be regulated spatially with Cre-recombination. Read the paper in Frontiers in Molecular...single step gRNA multiplexing system that generates vectors containing high numbers of gRNAs in one overnight...overnight reaction. STAgR is highly customizable as vector backbones, and gRNA structure, numbers, and promoters...Read the paper in Science Find the anti-CRISPR expression plasmids CRISPR-based method for cell lineage...Averof lab. It’s an alternative to the conventional Cre or Flp recombinases used to excise markers. It has...Cas12a inhibitors Using a bioinformatics and a screening approach, the Jennifer Doudna lab identified three...

  39. Custom CRISPR Screens & the Green Listed Software

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    Blog Post
    ...custom CRISPR screens Differentially expressed genes  As the costs for global gene expression analyses like...transcriptomics data. Custom CRISPR screens designed to target differentially expressed genes can be the next stage...These targeted screens can allow a researcher to test which of the differentially expressed genes found ...large extent involve receptors expressed on the cell surface. Thus, a screen can be used to target surface...example, you could design a screen targeting all differentially expressed genes that are known drug targets... your vector of choice using the adapter sequences that you have specified.   Run your screen! Final...and Feng Zhang. "Improved vectors and genome-wide libraries for CRISPR screening." Nature methods 11.8 (...

  40. Open Resources and Plasmid Tools For Studying C. elegans

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    ... 2016 created cGAL, a GAL-4 based bipartite expression system that can regulate gene expression in C.elegans...and plasmid assembly line for the creation of CRISPR targeting vectors that each contains a guide RNA and...CRISPR techniques and new tools for conditional expression and degradation. These workshops not only highlighted...elegans open resources The C. elegans Neuronal Gene Expression Map & Network (CeNGEN) The CeNGEN goal is to ...to establish a comprehensive gene expression atlas of every C.elegans  neuron. Scientists are currently...C.elegans. In this system a promoter controls the expression of a GAL-4 driver that can bind to activation...activation sequences upstream of genes to stimulate expression (Wang et al., 2016). In 2018, the lab expanded ...
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