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  1. PiggyBac-ing Through the Genome Editing Field

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    Blog Post
    ...plasmids with the piggyBac™ transposon. These plasmids, when combined with a source of piggyBac™ transposase...genome. We encourage you to deposit your piggyBac™ transposon vectors with us to help us expand this useful...Goodwin from Transposagen What is the piggyBac™ transposon? The piggyBac™ transposon was originally identified... transposons, piggyBac™ has two components, a transposon and a transposase. The piggyBac™ transposase ...excision-only piggyBac™ transposase. Image courtesy of Transposagen.   How does piggyBac™ enhance gene...technology with the piggyBac™ system, these challenges can be considerably reduced. The piggyBac™ transposon ...removed using the piggyBac™ transposase. This can be best achieved by utilizing a piggyBac™ transposon in...
  2. Plasmids 101: Using Transposons in the Lab

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    Blog Post
    ...Sleeping Beauty plasmids  piggyBac Although its name suggests otherwise, piggyBac was discovered in the cabbage... Beauty, piggyBac has an integration efficiency in human cells similar to retroviral vectors. The hyperactive...compared to the codon-optimized wild-type piggyBac transposase.  piggyBac plasmids    Tol2  Tol2 was the first...with Sleeping Beauty and piggyBac, which is a big advantage over viral vectors (~5 kB payload for AAV and...in viral vectors, but easily fit in a TE. Transposons are also less likely than viral vectors to induce...transposon’s LTRs, similar to when packaging viral vectors.   Let’s take a closer look at three different ...adapted for use as research tools: Sleeping beauty, PiggyBac, and Tol2.  Sleeping beauty Sleeping Beauty is...
  3. Hot Plasmids - August 2020

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    Blog Post
    ... Institute developed fluorescently labeled piggyBac vectors for uniform and sustained expression of single...both dCas9 and gRNAs. The multi-gRNA piggyBac vector integrates into genomic DNA with the help of...pMAK backbone by modifying pMAL-T-Avi-His/BirA, a vector commonly used for in vivo biotinylation via BirA... by a doxycycline inducible promoter. The gRNA piggyBac plasmids include a mRFP reporter. With this system...transcript and protein levels. Plasmids for multiplexed piggyBac gRNA delivery and the all-in-one dCas9-KRAB and...found at Addgene. Check out these plasmids for piggyBac gRNA delivery and the all-in-one dCas9 targeting...
  4. What's New in CRISPR - March 2020

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    Blog Post
    ...’s lab developed new fluorescently labeled piggyBac vectors to deliver uniform and sustained expression... Find the CRISPRi plasmids A multiplexed gRNA piggyBac transposon system for human pluripotent stem cells...
  5. Delivery Methods for Generating iPSCs

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    Blog Post
    ...residual expression piggyBac Excisible Medium Medium Generates transgene-free and vector-free cells Genome...so either a single polycistronic vector or multiple single-gene vectors can be used to package reprogramming...reprogramming vectors available from Addgene. Another drawback is that lentiviral vectors insert in different...Cre-lox lentiviral vectors: This approach is the same as using a standard lentiviral vector, but transient...Non-integrating iPSC delivery methods Viral Vectors Adenovirus: Adenoviral vectors infect dividing and nondividing...reprogramming factors: Mouse Expression Vectors Human Expression Vectors Carey et al Hockemeyer et al Brambrink...Non-integrating Low Medium Generates transgene-free and vector-free cells No genomic integration Inefficient...
  6. Plasmids 101: The protein expression toolbox

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    Blog Post
    ...out Addgene's Lentiviral dCas9 SAM vector and Piggybac dCas9 SAM vector! On/off control all the time If ...knock out plasmids? Check out our Cre vector and Murine LoxP vector.   Turn it off for a little while Transient...? Check out our Retro expression vector and Lenti expression vector. Turn it on for just a little while...Addgene's Lentiviral dTAG cloning vector and SMASh degron tagging vector! Destabilizing domains Destabilizing...Lentiviral Tet-on and AAVS1 targeted Tet-on transgene vector. Degron tags Tagging relevant proteins with a degron...levels. Looking for a DD? Check out our DD-Cas9 vector! The price of tunable control These systems can...
  7. 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...
  8. Cpf1 Update: Comparison to Cas9 and NgAgo

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    Blog Post
    ...deliver in low-capacity vectors like AAV. AAV (adeno-associated viral vectors) are commonly used for in...CRISPR Competitor NgAgo Learn How You Can Use piggyBac Vectors in Genome Editing Resources on Addgene.org...preferential infection of certain tissues. However, these vectors have a low genetic capacity of ~4.5 kb. At 4.1 ...previously used SaCas9 (1053 aa/3.2 kb) with AAV vectors for efficient postnatal editing. LbCpf1 and AsCpf1...SpCas9. These nucleases should fit well into AAV vectors, especially given that Cpf1 gRNAs are also shorter...
  9. 15 Hot Plasmids from 2017

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    Blog Post
    ...reprogramming discrepancies. The polycistronic piggyBac transposon vectors tested in this work are available at ...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...lead to comparable experimental results. Using PiggyBac transposons, the Woltjen Lab compared different...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...
  10. Plasmids for Stem Cell Research

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    Collection
    ...lentiviral vector for the expression of human Oct4, Sox2, and Klf4 Polycistronic lentiviral vector for "hit...25. 126(4):663-76. Yamanaka piggyBac Mouse Doxycycline-inducible piggyBac transposon reprogramming system...2011 Mar;21(3):518-29. Cheng piggyBac Mouse Doxycycline-inducible piggyBac transposon reprogramming system...Lentivirus Human Single polycistronic lentiviral vector for the expression of human Oct4, Klf4, Sox2, and...polycistronic, doxycycline-inducible lentiviral vector for the expression of human Oct4, Klf4, c-Myc, ...transcriptional activating fusions in single lentiviral vectors for human iPSC generation OCT4 and SOX2 Work as...Polycistronic, doxycycline-inducible lentiviral vectors for the expression of mouse Oct4, Klf4, c-Myc, ...
  11. CRISPR Plasmids - Prime Edit

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    Collection
    ...Publication Empty Prime Editing gRNA Vectors A selection of empty gRNA vectors suitable for prime editing are...-FokI Screen Pooled Libraries gRNAs Empty gRNA Vectors gRNAs By Species Mammalian Bacteria Drosophila ... table below. Use the search bar to find a gRNA vector based on expression system, promoter, the type ...Tilmann Weber 173220 pPBT-peRNA_GG-Puro Mammalian, piggyBac hU6 pegRNA BsmBI No Puromycin Jacob Giehm Mikkelsen...Mikkelsen 173222 pPBT-PE2-PuroTK-pegRNA_GG Mammalian, piggyBac hU6 pegRNA BsmBI Yes (Cas9 H840A + MMLV RT) PuroTK...
  12. 27 Hot Plasmids from 2016

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    Blog Post
    ...expression vectors with various promoters suitable for in vivo expression, and/or produce vectors to make...prototrophic strains. The vectors were based on the original EasyClone vectors (Jensen et al 2013) allow...donor vectors containing mAID and either Neo or Hygro resistance markers with CRISPR/Cas vectors targeting...uses the piggybac transposase to integrate dCas9-VPR into the genome of target cells. The piggybac system... interest and cloned into provided destination vectors using low-cost, one-pot golden gate cloning that... Jacks Lab constructed several GMAP-compatible vectors for a wide variety of biological applications. ... problem, Csaba Pál’s lab has created a set of vectors that allows one to use the MAGE method in unmodified...
  13. Neurodegeneration Research Collection

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    Collection
    ...motor neuron-specific reporter and a PAX7 inducible vector . Harley et al. Cell Rep. 2023 Dec 26. Induce CNS... toolbox . Lam et al. bioRxiv. 2022 Dec 2. Use PiggyBac plasmids with tet-inducible expression of transcription...with rapid delivery. Includes antibodies, viral vectors, animal models, purified protein, and more. Plasmids...
  14. Neurodegeneration Plasmid Collection

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    Collection
    ...19 James Trimmer 129409 Slc1a3-CreERT2 Targeting Vector SLC1A3 PGK Episodic ataxia Walker Jackson 129410...CMV Spinocerebellar Ataxia Richard Youle 176485 Piggybac-TetO-hNurr1-blast NR4A2 TetO Parkinson's Marius...ApoE4 APOE CMV Alzheimer's Bradley Hyman 197908 piggybac-EF1-halo-mTBP TBP Halo EF1a Parkinson's James ...GST-TEV-TBK1 TBK1 polH ALS Sascha Martens 209078 piggyBac-rtTA (4th_Gen)-SNCA(3K)-sfGFP-IRES-NGN2-puro (...SNCA GFP TRE Parkinson's Vikram Khurana 209080 piggyBac-rtTA (4th_Gen)-SNCA(A53T)-sfGFP-IRES-NGN2-puro...SNCA GFP TRE Parkinson's Vikram Khurana 209081 piggyBac-rtTA (4th_Gen)-SNCA(A53T-DNAC)-sfGFP-IRES-NGN2...SNCA Avi CMV Parkinson's Vikram Khurana 209085 piggyBac_rtTA (4th_Gen)_SNCA_IRES_NGN2-2A-PURO_Synapsin-BirA...
  15. CRISPR Guide

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    Collection
    ...format into lentiviral transfer vectors (Figure 8B). Libraries come in 1-vector systems, in which Cas9 is included...targeting in the double digits. These multiplex gRNA vectors can be used to knock out, activate, or repress ...multiplexing . Browse Plasmids: Multiplex gRNA Vectors Engineering the Cas9 Endonuclease CRISPR specificity...enzymes. CRISPR Transposases Transposon systems like piggyBac or Tn7 allow for large scale edits by integrating...heterogeneous population of lentiviral transfer vectors. In most cases, each plasmid contains an individual... included in the gRNA-containing plasmid, or 2-vector systems, in which Cas9 must be delivered separately...experimental cell population (Figure 8E). In a 2-vector system, you’ll need to either co-infect with a ...
  16. CRISPR Guide

    Type
    Guide
    ...format into lentiviral transfer vectors (Figure 8B). Libraries come in 1-vector systems, in which Cas9 is included...targeting in the double digits. These multiplex gRNA vectors can be used to knock out, activate, or repress ...multiplexing . Browse Plasmids: Multiplex gRNA Vectors Engineering the Cas9 Endonuclease CRISPR specificity...enzymes. CRISPR Transposases Transposon systems like piggyBac or Tn7 allow for large scale edits by integrating...heterogeneous population of lentiviral transfer vectors. In most cases, each plasmid contains an individual... included in the gRNA-containing plasmid, or 2-vector systems, in which Cas9 must be delivered separately...experimental cell population (Figure 8E). In a 2-vector system, you’ll need to either co-infect with a ...
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