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We narrowed to 330 results for: NUC

Showing: 61 - 80 of 330 results

  1. SARS-CoV-2/COVID-19 Detection Methods Based on CRISPR/Cas

    Type
    Blog Post
    Published
    May 5, 2020, 1:15 p.m.
    ...OO, Zhang F (2019) SHERLOCK: nucleic acid detection with CRISPR nucleases. Nature Protocols 14:2986–3012...the DIY-Bio movement. Rapid, field-deployable nucleobase detection and identification using FnCas9 (FELUDA...positions of these mismatches. To detect SARS-CoV-2 nucleic acids, the target sequence is amplified by PCR/...FnCas9, this is also a good way to diagnose single nucleotide variations. Also, FnCas9 cleaves in a range of..., … Maiti, S. (2020). Rapid, field-deployable nucleobase detection and identification using FnCas9. https...

  2. Comparing Cas9 to NgAgo: Can the Argonautes Best CRISPR?

    Type
    Blog Post
    Published
    June 9, 2016, 2:30 p.m.
    ...Pyrococcus furiosus is a DNA-guided nuclease that targets cognate DNA.” Nucleic Acids Res. 43(10) (2015):5120...user-friendly, since they can be ordered as oligonucleotides and should be stable in the cell. gDNA guides...a different approach, randomly removing 1-20 nucleotides from the cleavage site specified by the gDNA.... While this fidelity would prevent other small nuclear DNAs from binding NgAgo, it might make it difficult...

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

    Type
    Blog Post
    Published
    Sept. 26, 2024, 1:15 p.m.
    ... how far the T4 exonuclease proceeds: at the first occurrence of the chosen nucleotide, T4 will switch...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...overhangs must not contain the dNTP present in the exonuclease reaction or polymerization will occur at that...

  4. Why and how to start studying chromatin (with Addgene's help)

    Type
    Blog Post
    Published
    June 12, 2018, 11:40 a.m.
    ...Certainly referring to any DNA within the context of a nucleus as “linear” is a misnomer. The DNA packed into ... may be looking at chromatin at the level of nucleosomes, loops, chromosomal domains, or how stretches... research. DNA Cross-Bridging Shapes a Single Nucleus from a Set of Mitotic Chromosomes. Samwer M, Schneider...2):197-211. DNA Cross-Bridging Shapes a Single Nucleus from a Set of Mitotic Chromosomes. Samwer M, Schneider... 25. 95(5):625-36. An architectural role for a nuclear noncoding RNA: NEAT1 RNA is essential for the structure...

  5. Kiran Musunuru on the Newest TALEN Genome-Editing System

    Type
    Blog Post
    Published
    Dec. 10, 2013, 3:13 p.m.
    ...cells: a transcription activator-like effector nuclease (TALEN) kit consisting of 834 plasmids. The researchers...proteins that comprise a DNA binding domain and an endonuclease domain. What’s distinctive is that the DNA-binding... rest of the protein in place including FokI endonuclease domain as well as fluorescent proteins used ...first really popular tool was the zinc finger nucleases. Unfortunately, it is time consuming and difficult...without a lot of expertise to create zinc finger nucleases that will bind desired sequences. TALENs are much...

  6. CRISPR 101: Cas9 vs. The Other Cas(s)

    Type
    Blog Post
    Published
    Dec. 8, 2022, 2:15 p.m.
    ...We are all oh so familiar with the nuclease Cas9, but how does it stack up to the new Cas enzymes on ...without shortly thereafter hearing ‘Cas9’. The Cas9 nuclease is a genome engineering powerhouse, and for good... often hard to detect variants such as single nucleotide polymorphisms. Fast facts – PAM requirement:...it – literally. Cas13 Cas13 is an RNA-guided nuclease, however, unlike Cas9, it targets RNA instead ...., Omar, et. al. Cpf1 is a single RNA-guided endonuclease of a class 2 CRISPR-Cas system. Cell, 163(3)...

  7. Progress Towards a PAM-Free CRISPR

    Type
    Blog Post
    Published
    March 7, 2024, 2:39 p.m.
    ...anymore. Say hello to PAM-free and PAM-flexible nucleases! In this blog, we will review Cas enzymes, which...sequence, that of SpCas9, is NGG. Assuming all nucleotides are represented equally, that’s only a 1/16 chance... NG Cas9 NG - SpG Cas9  NG  - SpRY nuclease  C to T Base Editor SpRY A to G Base Editor SpRY...  Nishimasu, H., et al.,Engineered CRISPR-Cas9 nuclease with expanded targeting space. Science 361,1259...compatibility and editing activities of Cas9 variants. Nucleic Acids Res. 2021 Sep 7;49(15):8785-8795. doi: 10.1093...

  8. Degrading DNA with Cascade-Cas3

    Type
    Blog Post
    Published
    Feb. 11, 2025, 2:15 p.m.
    ...chop DNA. Cas3, however, has two jobs, one as a nuclease and one as a helicase. Following recruitment by...initial steps, Cas3 can perform its helicase and nuclease functions simultaneously, by continuously unraveling... into think it’s more powerful than it is. The nuclease domain of Cas3 is considered weak, especially ...However, by continually passing the ssDNA past the nuclease domain, Cas3 is able to repeatedly cut the DNA...Repetitive DNA reeling by the Cascade-Cas3 complex in nucleotide unwinding steps. Molecular Cell, 70(3), 385-394...

  9. Adenoviral Delivery of CRISPR/Cas9 Aims to Expand Genome Editing to Primary Cells

    Type
    Blog Post
    Published
    Dec. 10, 2020, 1 p.m.
    ...that AdV-mediated transduction of gRNA:Cas9 ribonucleoprotein complexes into transformed and non-transformed...chromosomal region. The CRISPR/Cas9-derived RNA-guided nuclease-induced gene disruption frequencies in the various...Methods the same year that delivering RNA-guided nucleases or TALENs together with AdV donor DNA leads to...DNA for accurate genome editing with engineered nucleases. Nat Methods 11:1051–1057 . https://doi.org/10.1038...Adenoviral vector delivery of RNA-guided CRISPR/Cas9 nuclease complexes induces targeted mutagenesis in a diverse...

  10. Polymerase Chain Reaction Overview and Applications

    Type
    Blog Post
    Published
    Aug. 10, 2021, 1:15 p.m.
    ...addition to Taq DNA polymerase, PCR requires free nucleotides (dNTPs), template DNA to amplify from and unique...polymerases require an existing strand of DNA to add nucleotides to.  Using these reagents and a series of heating...method that provides ultrasensitive and absolute nucleotide concentration, unlike qPCR where results can ... ddPCR Determine ultrasensitive and absolute nucleotide concentration of starting material AAV Titrations... References Shen C-H (2019) Amplification of Nucleic Acids. In: Diagnostic Molecular Biology. Elsevier...

  11. Optogenetics + CRISPR, Using Light to Control Genome Editing

    Type
    Blog Post
    Published
    Sept. 3, 2020, 12:15 p.m.
    ...(730 nm). This nucleotide triggers the translocation of the protein BldD to the nucleus. To couple this...system is unique in that it utilizes a “split” nuclease - the authors fragmented Cas9 into N-terminal ...al., 2016 CRISPR-plus Photocleavable ssDNA oligonucleotides Cas9, photocaged-gRNA Gene editing Manna...transcription Zhou et al., 2020   NPOM-caged nucleobases Cas9, photocaged-gRNA Gene editing For more...temperature- and light-switchable Cas9 variants. Nucleic Acids Research 44: . https://doi.org/10.1093/nar...

  12. QC Sequencing Technologies at Addgene

    Type
    Blog Post
    Published
    July 15, 2025, 1:15 p.m.
    ... bases (A, C, T, or G) is supplied as a dideoxyribonucleoside triphosphate (ddNTP) so that after its addition... reads a fluorescent label on the terminating nucleotide and records it in a “trace” file to determine...through the pore via motor proteins. When each nucleotide passes through the nanopore, the electrical current.... Base-calling software can then identify the nucleotide that passed through based on these characteristic...pass through our QC process if there is a 100% nucleotide match to base identity and plasmid length. Any...

  13. Four Base Editing Reporters to Monitor and Enrich Editing in Real-time

    Type
    Blog Post
    Published
    July 7, 2020, 1:15 p.m.
    .... APOBEC-Cas9n-UGI targets its preferred trinucleotide motifs (5’-TCA-to-TUA) found within the 43 base-pair...Cas9-mediated Genome Editing. Molecular Therapy - Nucleic Acids 5:e334 . https://doi.org/10.1038/mtna.2016.48... to identify and enrich base editing activity. Nucleic Acids Research 48:2841–2852 . https://doi.org/10.1093...APOBEC–Cas9 or cleavage by Cas9 in living cells. Nucleic Acids Research 46:e84–e84 . https://doi.org/10.1093... for editing enrichment (TREE) in human cells. Nucleic Acids Research 47:e120–e120 . https://doi.org/10.1093...

  14. Plasmids 101: Biotinylation

    Type
    Blog Post
    Published
    Nov. 15, 2018, 1:50 p.m.
    ...to an oligonucleotide depending on your downstream application. Biotin labelled oligonucleotides can be...that contain both AviTag and BirA at Addgene! Oligonucleotide labeling Biotin can be added to either terminus... becomes reactive and forms a link with nearby nucleic acids (McInnes et al., 1990). Photobiotinylation...specific dsDNA, ssDNA or RNA–protein interactions.” Nucleic Acids Research, 23.18 (1995): 3792–3793. PubMed...

  15. CRISPR 101: Engineering the Plant Genome Using CRISPR/Cas9

    Type
    Blog Post
    Published
    Oct. 11, 2016, 2:30 p.m.
    ...the gRNA (composed of a ~20 nucleotide targeting sequence and ~75 nucleotide scaffold sequence) is consistent...commonly tagged with a nuclear localization sequence to enhance targeting to the nucleus, and several codon...plant species or cell type (Belhaj et al., 2013). Nuclease dead Cas9 (dCas9) based activators (such as dCas9...

  16. Tips for a 1st Time CRISPR User (by a 1st Time CRISPR User)

    Type
    Blog Post
    Published
    March 7, 2017, 3:30 p.m.
    ...various MOIs. The presence of the 428 bp Surveyor nuclease cleavage product indicates a gene edit. GFP gRNA-transduced...The basis of the Surveyor assay is that Surveyor nuclease cleaves double stranded DNA (dsDNA) at basepair...mismatches at the site of the edit. Using Surveyor nuclease followed by gel electrophoresis, these mismatches...Before digesting the DNA duplexes with Surveyor nuclease, save some of the duplex and run it as an undigested...BRAF sites were edited in A549 cells. Surveyor nuclease was used to digest (+) mismatches in DNA duplexes...

  17. Quick, Versatile Plant Transgenesis with GreenGate Plasmids

    Type
    Blog Post
    Published
    Jan. 23, 2014, 3:23 p.m.
    ...GreenGate are type IIS restriction endonucleases. They are like other endonucleases except that they cut at arbitrary...be very careful with the type IIS restriction endonuclease of choice, BsaI, which can be very sensitive... They finally traced the problem back to the endonuclease. “It’s just a sensitive protein,” he says. “...

  18. What's New in CRISPR - December 2019

    Type
    Blog Post
    Published
    Dec. 17, 2019, 2:37 p.m.
    ...tagging Lentiviral capsid-based Cas9/gRNA ribonucleoprotein delivery Cytosine deaminase for RNA editing...bioRxiv Find the ORANGE toolkit   Cas9/gRNA ribonucleoprotein delivery by lentiviral capsid-based bionanoparticle...bionanoparticle system for packaging Cas9/gRNA ribonucleoproteins for transient expression. They replaced the...a lentivirus-like particle. Read the paper in Nucleic Acids Research Find the gRNA and packaging plasmids...

  19. CRISPR 101: Cytosine Transversion Editors

    Type
    Blog Post
    Published
    March 25, 2025, 1:15 p.m.
    ...Note: hypoxanthine (shown) is the nucleobase component of the nucleoside inosine. Created with BioRender.com...improved the CGBE: DNA polymerase D2 (POLD2), exonuclease 1 (EXO1), and RNA binding motif protein X-linked...Repair A cellular process in which incorrect nucleobases are removed from the backbone, creating abasic...further repair.   Abasic site A DNA site with the nucleobase removed; also called an apurinic / apyrimidinic...

  20. CASTing Off for New Shores in Human Genome Editing

    Type
    Blog Post
    Published
    June 10, 2025, 1:15 p.m.
    ...complex minus its nuclease component, Cas3, while Type V-K CAST systems use a nuclease-deficient Cas12k..., Tn7-like transposons captured and repurposed nuclease-deficient CRISPR-Cas systems to help them spread...Gelsinger et al., 2024). In either type of CAST, the nuclease-deficient CRISPR-Cas system identifies and binds...
Showing: 61 - 80 of 330 results