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We narrowed to 103 results for: EXO

Showing: 1 - 20 of 103 results

  1. Improving Transduction Efficiency with Exosome AAVs

    Type
    Blog Post
    ... vectors and exosome was given the name vexosomes (vector-exosomes), exosome AAVs, or exo-AAVs (Figure...efficacy of AAV vectors. How are exosome AAVs produced? Purifying exo-AAVs is actually a bit simpler than...behind these improvements. What have exosome AAVs achieved so far? Exo-AAVs have improved transduction profiles...as retina or glioblastoma). They also found that exo-AAVs were more resistant to neutralizing anti-AAV...The current purification protocols for isolating exo-AAVs rely on differential centrifugation steps to...to separate cell culture supernatant containing exo-AVVs from producer cells, and then to enrich the microvesicles...containing AAVs (Figure 2). Since the development of the exo-AAV isolation protocol by Maguire and colleagues,...

  2. Lambda Red: A Homologous Recombination-based Technique for Genetic Engineering

    Type
    Blog Post
    ...introduced into the E. coli. Exo: Exo is a 5’→3’ dsDNA-dependent exonuclease.  Exo will degrade linear dsDNA...recombineering system has three components (Figure 1): 1) Exo, 2) Beta, and 3) Gam. All three are required for ...degraded. Beta: Beta protects the ssDNA created by Exo and promotes its annealing to a complementary ssDNA...system).  In some of these strains, expression of exo, beta and gam is tightly regulated by the endogenous...repressor to re-nature and again repress expression of exo, beta and gam.  A major advantage of using this method...

  3. Open Enzyme Collection

    Type
    Collection
    ...165522 pOpen-K12polLF (exo-) DNA Polymerase I, Large (Klenow) Fragment (3'-5' exo-) 165512 pOpen-THEAQpolA... Polymerase IV 165523 pOpen-Tlipol (exo-) Tli DNA polymerase (exo-) 165524 pOpen-T4gene43 T4 DNA Polymerase...pOpen-P45 P45 165572 pOpen-GBDpol (exo) Pyrococcus sp. Heat-Stable (exo-) DNA Polymerase 165573 pOpen-phi29pol... pOpen-Exonuclease III E. coli E. coli Exonuclease III 165559 pOpen-T5gene12/13 T5 Exonuclease 165560 ...

  5. Ligation Independent Cloning

    Type
    Protocol
    ...without ligation. Because of its dual polymerase/exonuclease functions, T4 DNA polymerase can create overhangs... which a single dNTP is included to stop the exonuclease function of the enzyme and shift its activity...polymerase protocol), causing the enzyme to perform exonuclease activity until the first "G" in the sequence.... the polymerase reaction is favored over the exonuclease reaction, the polymerase will add back the guanosine... proceeding, as they will interfere with the exonuclease activity of T4 Pol in the following step. This...

  6. Gibson Assembly Protocol

    Type
    Protocol
    ...specific and unique function for the reaction: T5 Exonuclease - creates single-strand DNA 3’ overhangs by chewing...the ssDNA-specific endonuclease activity of T5 Exonuclease and furthermore reduces secondary structure of...

  7. Treating Muscular Dystrophy with CRISPR Gene Editing

    Type
    Blog Post
    ...trials have used oligonucleotide exon skipping (OEN) to remove mutated exons from the dystrophin transcript...nucleases to remove exon 51 in DMD patient myoblasts. They observed a 13% removal rate of exon 51, which resulted... they used CRISPR with two gRNAs to delete exon 51 or exons 45-55 in patient myoblasts; when injected ... that exon skipping therapies would be applicable to 80% of DMD patients. Figure 1: Exon skipping...each chose to try exon skipping in DMD mice, which have a mutation in dystrophin exon 23. Long et al. used...mutations in human-relevant exons 45-55, rather than in the traditional exon 23-mutated mouse model. Potential... editing exonic regions comes with much more risk than the intronic editing used in the exon skipping ...

  8. Plasmids 101: Knockout/Knock-In Plasmids

    Type
    Blog Post
    ...cell line with the floxed exon. You can then conditionally remove this exon (and hence knock out the gene...walk through an experiment designed to knock out exon 2 of a given gene.   Figure 1: A knockout...shown in Figure 1, large sequences corresponding to exons 1 and 3 of the targeted gene have been cloned into...construct to your target cells. After recombination, exon 2 of the targeted gene will be removed from the ...to insert, or knock-in, any gene, tag, or mutated exon into the genome. For this purpose, the sequence ...with the Neomycin resistance (NeoR) gene between exon 1 and 3 of the targeted gene. Upon recombination...recombination, the GFP/NeoR cassette is inserted in place of exon 2. Thus, the targeted gene is disrupted (knocked...

  9. Deep Mutational Scanning with One Pot Saturation Mutagenesis

    Type
    Blog Post
    ...UDG) and Exonuclease III (ExoIII) (step 4). UDG recognizes uracils that are in DNA and ExoIII recognizes... strands of DNA. Treating the plasmid with ExoIII and ExoI then fully degrades the nicked strand. Synthesize... step 1. Then this strand is degraded with ExoIII and ExoI. Synthesize 2nd mutant strand: The second mutant...digest and the whole plasmid will be degraded by ExoIII. See the below figure for an example like this....

  10. The Breast Cancer Microenvironment: A Tumor’s Backstage Team

    Type
    Blog Post
    ...that, let’s take a look at exosomes. Proficient in transportation, exosomes are tools of communication...after injecting breast cancer cells into mice, exosomes expressed specific integrins (cell-adhesion proteins...proteins) targeting different organs. This means that exosomes not only promote microenvironment’s malignancy...also incite different organs to become malignant. Exosomes also contain miRNA (micro RNA) and other molecules...epigenetic changes in targeted cells. In this case, exosomes may transport miRNA from drug-resistant cells ...PMID: 22439926. Hoshino, Ayuko, et al. "Tumour exosome integrins determine organotropic metastasis." Nature...

  11. Cas13d: Small RNA-targeting CRISPR enzymes for transcriptome engineering

    Type
    Blog Post
    ...to manipulate RNA species through exon skipping. Previous CRISPR exon-skipping approaches used two guide...guide RNAs to remove a given exon from the genome, and showed success in models of muscular dystrophy. In...delivering dCasRx and a 3-spacer array targeting the MAPT exon 10 splice acceptor and two putative splice enhancers...to iPS-derived cortical neurons, dCasRx-mediated exon skipping improved the ratio of pathogenic to non-pathogenic...

  12. Fluorescent Protein Guide: Biosensors

    Type
    Collection
    ...pH-sensitive red fluorescent protein for imaging of exo- and endocytosis. J Cell Biol. 2014 Nov 10;207(3)...Endo/Exocytosis Superecliptic pHluorin fused to glutamate receptor Glutamate receptor exocytosis and spine...2006 Feb 15. 26(7):2000-9. Robert Malinow Endo/Exocytosis Synaptophysin-pHluorin Clathrin-mediated endocytosis... 2006 Sep 21. 51(6):773-86. Leon Lagnado Endo/Exocytosis Synaptophysin with tandem repeats of pHluorin...Prescott pH Synaptic vesicle pH sensors for imaging exocytosis and endocytosis (pHuji). Semisynthetic fluorescent...fluorescent pH sensors for imaging exocytosis and endocytosis. Nat Commun. 2017 Nov 10;8(1):1412. Justin Taraska...

  13. Plasmids 101: Sequence and Ligation Independent Cloning (SLIC)

    Type
    Blog Post
    ...The key is T4 DNA polymerase, which has 3′→5′ exonuclease activity. T4’s polymerase activity can be thwarted...available to polymerize, allowing the slower exonuclease activity to proceed. By including only one type...researcher can decide exactly how far the T4 exonuclease proceeds: at the first occurrence of the chosen...overhangs must not contain the dNTP present in the exonuclease reaction or polymerization will occur at that...

  14. Plasmids 101: The protein expression toolbox

    Type
    Blog Post
    ...the control happening? Is the regulation of an exogenously introduced gene or over the native locus? Let...knocked in flanking a gene of interest (or a single exon) and everything in between those sites can be excised... systems work by tagging the endogenously or exogenously expressed protein with the relevant tag and then... These systems can be easily implemented for exogenously introduced proteins, as many of them just need...

  15. Plasmids 101: Gibson Assembly and Other Long-Homology Based Cloning Methods

    Type
    Blog Post
    ...First, it uses a dedicated 5’ exonuclease instead of using the exonuclease feature of T4 DNA polymerase... which contains three different enzymes: an exonuclease, which chews back the 5’ ends of the fragment...over 200 nucleotides. This may be because the exonuclease could chew through an entire fragment shorter...

  16. Plasmids 101: Common Lab E. coli Strains

    Type
    Blog Post
    ...recombination, increases plasmid stability recBCD Exonuclease V activity abolished Increased plasmid stability...thi1 Mutation in thiamine metabolism Requires exogenous thiamine for growth ara disruption of arabinose...β-isopropyl malate dehydrogenase inactivated Requires exogenous leucine source for growth proAB mutation in ...in proline biosynthesis pathway Requires exogenous proline source for growth rpsL Mutation in subunit...

  17. RNA Interference in Plant Biology: New Tools for an Old Favorite

    Type
    Blog Post
    ...result, gene silencing can be easily induced by exogenous application of long or short dsRNA. However, it...unmatched—unlike CRISPR experiments that require exogenous protein, successful induction of RNAi only requires... can maintain partial function due to the use of exon skipping, alternative start codons, and/or the phenomenon...microRNAs found in the genome, whereas shRNAs are exogenously derived. Importantly, instead of the assortment...

  18. Plasmids 101: Gene, enhancer, and promoter trapping

    Type
    Blog Post
    ...promoter-less reporter which needs to integrate within an exon of a gene to generate a transcriptional fusion. ...do not need to be integrated within an intron or exon of a gene, and thus they do not disrupt the gene...While this system does not require insertion into an exon, like promoter traps, it will still likely disrupt...

  19. Hot Plasmids - October 2020

    Type
    Blog Post
    ...paralogs. The exon-deletion hybrid gRNA library deletes human alternative cassette exons and disrupts ...disrupts genes with alternative exons. Get the library from Addgene New from the viral service By Jason Nasse...

  20. CRISPR 101: Targeting Non-Coding RNAs with CRISPR/Cas9

    Type
    Blog Post
    ...mutate the transcription start site, splice sites, exons, and/or promoters to knock out lncRNA function (...result in the retention of introns or the removal of exons. Disrupting promoter or transcription start site...(B) Mutations of splice sites. (C) Removal of an exon or a large genomic fragment. (D) Knock-in: insertion...
Showing: 1 - 20 of 103 results