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Showing: 1 - 11 of 11 results
  1. 15 Years of Addgene: The Top 15 Plasmids

    Type
    Blog Post
    Published
    Jan. 8, 2019, 1:54 p.m.
    ...  Find the scrambled shRNA vector. pLKO.1 - TRC cloning vector - This vector from the David Root lab is...lentiGuide-Puro. pSpCas9n(BB)-2A-GFP (PX461) - Similar to pSpCas9(BB)-2A-Puro (PX458), pSpCas9n(BB)-2A-GFP (PX461...sequence (pSpCas9(BB)). The 2A-GFP allows for screening of transfected cells. Find pSpCas9(BB)-2A-GFP (PX458...Find pSpCas9n(BB)-2A-GFP (PX461). pSpCas9(BB)-2A-Puro (PX459) - This mammalian expression vector encodes...top requested plasmid of 2017. Find pMD2.G. pSpCas9(BB)-2A-GFP (PX458) - This plasmid from the Zhang lab...from the Zhang lab is a mammalian vector that expresses both SpCas9 and your gRNAs and was also the top...opening our UK office in 2014, to starting our viral vector service in 2016...we’re excited to help scientists...
  2. Multiplex Genome Editing with CRISPR-Cpf1

    Type
    Blog Post
    Published
    May 9, 2017, 2:12 p.m.
    ...expression vectors which are often more difficult to transfect or package into viral vectors. Cas9 multiplexing...array plus a GFP tag. Four weeks post-infection, there was strong expression of Cpf1 and GFP in the targeted...Cassettes lentiviral transduction One vector expresses Cas9, eGFP, and up to 4 gRNAs Each gRNA requires... 1) Most rely on transfection of more than one vector to express the gRNAs and Cas9. Co-transfections...multiplexing vectors are larger because they require regulatory sequences to allow for expression of multiple...Yamamoto Lab Golden Gate Assembly transfection One vector expresses Cas9 and up to 7 gRNAs Each gRNA requires...plasmids resulting in smaller, more streamlined vectors which are easier to use across multiple expression...
  3. 22 Hot Plasmid Technologies from 2014

    Type
    Blog Post
    Published
    Jan. 6, 2015, 5:21 p.m.
    ...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... existing transposon and vector systems and creating an all-synthetic vector that included only the elements... into a destination vector in a predefined order. Finally, the destination vectors are used with Multisite... eight entry vectors can be used for each Multisite Gateway compatible destination vector for a maximum...step, compatible Level 0 vectors are directionally assembled into a Level 1 vector creating a single transcriptional...of interest into the pCoofy vectors, select the appropriate pair of vector and gene primers from Table...
  4. Your Top Requested Plasmid in 2016!

    Type
    Blog Post
    Published
    Jan. 11, 2017, 8:47 p.m.
    ...this vector contain GFP-tagged and un-tagged versions of SpCas9 and further variants replace SpCas9 with...plasmid from 2016 is an expression vector for CRISPR nuclease, SpCas9 from the Zhang Lab. It's name, though...versatile mammalian expression vector that you can use to express both SpCas9 and your gRNA in a variety ...versatile mammalian expression vector that you can use to express both SpCas9 and your gRNA of interest in...resistance gene as separate peptides. PX459 This is the vector backbone. This particular backbone can be used ... as is obvious from our most requested plasmid, SpCas9 is still going strong as the basis for many genome...further ado, the most request plasmid in 2016 was... pSpCas9(BB)-2A-Puro (PX459) V2.0       Similar to last...
  5. CRISPRainbow and Genome Visualization

    Type
    Blog Post
    Published
    Feb. 28, 2017, 3:30 p.m.
    ...expressed in a single vector, pCRISPRainbow-DONOR1, with dCas9 supplied by a separate vector. Figure ...signal to noise ratio of the system, they modified GFP-dCas9 and the gRNA scaffold to enhance complex assembly...the amount of background fluorescence from unbound GFP-dCas9. With these improvements, labeling efficiency...sequences, which recruit fluorescent proteins BFP, GFP, and RFP fused to the respective hairpin binding ...Fluorescent Proteins Control Protein Activity with GFP Resources on Asddgene.org Find Fluorescent Protein...create a colorful Cas9 toolbox, Ma et al. turned to SpCas9 and its orthologs NmCas9 and St1Cas9. Each ortholog...
  6. Hot Plasmids and Viral Preps - May 2021

    Type
    Blog Post
    Published
    May 18, 2021, 1:15 p.m.
    ...putative pre-synaptic terminals with one vector. AAV hSyn FLEx mGFP-2A-Synaptophysin-mRuby from Liqun Luo...  After validating this method through monitoring GFP degradation, this protein compartment was used to... a plant cloning kit that contains a series of vectors and DNA modules for assembly of CRISPR/Cas9 constructs...is available in serotype AAV1. The GCaMP8 viral vectors from the Janelia Farm GENIE project have arrived...arrived! We now offer several versions of GCaMP8 vectors in serotypes AAV1 and AAV5. AAV9 preps and more AAV1...Luo’s lab expresses membrane tethered GFP to enhance axonal labeling and synaptophysin-tagged mRuby to ...project. We now have over 40 INTRSECT 2.0 project vectors available as ready-to-use viral preps with more...
  7. Overcoming the AAV Size Limitation for CRISPR Delivery

    Type
    Blog Post
    Published
    Sept. 16, 2020, 1 p.m.
    ...Streptococcus pyogenes (SpCas9) and a gRNA together (~4.2 kb) into an AAV vector is challenging due to ...problems with Cas9 delivery. Adeno-associated viral vectors (AAV) are commonly used for in vivo gene delivery...packaged Cas9 and multiple gRNAs into separate AAV vectors, increasing overall packaging capacity but necessitating...markers were not increased compared to a control AAV-GFP. Not only did the AAV-SaCas9-sgRNA constructs mediate...genome engineering: An adeno-associated viral (AAV) vector toolbox. Biotechnology Journal 9:1402–1412 . https...Editing: Strategies and In Vivo Delivery by AAV Vectors. Cell 181:136–150 . https://doi.org/10.1016/j.cell...compared to the full-length SpCas9. In 2016, Chew et al. developed a split intein spCas9-AAV toolbox that retains...
  8. 15 Hot Plasmids from 2017

    Type
    Blog Post
    Published
    Jan. 9, 2018, 3:02 p.m.
    ...tags for mTagBFP, TagRFPt, EGFP, mVenus, mCerulean3, mKOFP2) and empty vectors that have standard cloning...neuronal tools to the expanding gateway compatible vectors currently shared amongst the zebrafish community...-positive assay) as well as stabilization (Nanog-GFP reporter activation and silencing of factor-linked...discrepancies. The polycistronic piggyBac transposon vectors tested in this work are available at Addgene. ...for the research community. You can find the AAV vectors for infecting GABA-ergic interneurons here. Dimidschstein...Very low off-target activity; protein purification vector  87439  pCMV-HF-BE3  HF-Cas9 (NGG)  Very low ...low off-target activity; mammalian expression vector Rees H, et al. Nature Communications. 2017. PubMed...
  9. With Our New Viral Service, We're Taking CRISPR Further!

    Type
    Blog Post
    Published
    Dec. 6, 2016, 3:30 p.m.
    ...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...Page Check Out Our CRISPR Guide Pages Find Viral Vectors ... including those targeting EGFP, BRAF, and MAP3K4. The gRNAs targeting EGFP can be used as positive controls... makes it unuseable. With the large size of the SpCas9 nuclease (~4.2 kb) this is a particularly prominent...Institute to provide lentiviruses for delivery of SpCas9 and Cas9 nickase for genome editing. You can also...
  10. Hot Plasmids and Viral Preps - July 2021

    Type
    Blog Post
    Published
    July 27, 2021, 1:15 p.m.
    ...inducible CRISPR inhibition (CRISPRi). A) Lentiviral vector design. B) Strategy for leak-free transcriptional...the new selection of ready-to-use AAV9 and AAVrg vectors. New red-shifted INTRSECT 2.0 calcium sensor available...your imaging experiments with the Cre-OFF/FLP-ON vector 137128-AAV8: Ef1a-Coff/Fon-sRGECO ...circularly permuted green fluorescent protein or cpGFP) to the petunia SL enzymatic receptor DAD2. SLs ...which modulates the fluorescence intensity of the cpGFP without altering that of the internal control (Fig...editing in moderate thermophiles.  These evolved SpCas9 variants have improved activity on NAG PAMS and...
  11. CRISPR 101: Epigenetics and Editing the Epigenome

    Type
    Blog Post
    Published
    June 24, 2020, 5:45 p.m.
    ...methylation and activate transcription. A lentiviral vector with the same modifier, Fuw-dCas9-Tet1CD, is available... targeted enhancers. Although the lab used this vector in chick embryos, it also functions in mammalian...and this fusion allows for packaging into viral vectors and targeted delivery into hard-to-manipulate cell...targeting to a wider set of genomic targets. These two vectors are part of the larger pMVP/pMAGIC cloning system...) Vlatka Zoldoš’ lab has deposited pdCas9-DNMT3A-EGFP and pdCas9-DNMT3A-PuroR for targeted cytosine methylation...methylation in mammalian cells. Co-expression markers EGFP and PuroR enable sorting and selection of transduced...Goodell's lab has deposited pcDNA3.1-dCas9-MQ1(Q147L)-EGFP, a fusion of dCas9 to a small DNA methyltransferase...
Showing: 1 - 11 of 11 results