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Enter a sequence to perform a BLAST-based search of Addgene plasmid sequence databases.

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We narrowed to 423 results for: NUC

Showing: 81 - 100 of 423 results

  1. Plasmids 101: Biotinylation

    Type
    Blog Post
    ...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...

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

    Type
    Blog Post
    ...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...

  3. Allen Institute for Cell Science Plasmid Collection

    Type
    Collection
    ...mTagRFP-T Nucleophosmin Nucleolus (granular component) 133963 UBTF-HaloTag AICSDP-80 HaloTag Nucleolar transcription...Golgi 109122 NPM1-mEGFP AICSDP-50 mEGFP Nucleophosmin Nucleolus (granular component) 109120 GJA1-mEGFP ...Cohesins 114407 NUP153-mEGFP AICSDP-50 mEGFP Nucleoporin Nup153 Nuclear pores 114410 FUS-mEGFP AICSDP-64 mEGFP...transcription factor UBF Nucleolus (fibrillar center) 133964 SON-mEGFP AICSDP-82 mEGFP SON Nuclear speckles 159744...Microtubules 87422 LMNB1-mEGFP AICSDP-10 mEGFP Lamin B1 Nuclear envelope 87423 TOMM20-mEGFP AICSDP-8 mEGFP Tom20...reticulum 87427 FBL-mEGFP AICSDP-13 mEGFP Fibrillarin Nucleolus (dense fibrillar component) 87428 MYH10-mEGFP ...114403 LMNB1-mTagRFP-T AICSDP-29 mTagRFPT Lamin B1 Nuclear envelope 114404 AAVS1-mEGFP (PGK) AICSDP-36 mEGFP...

  4. Open Enzyme Collection

    Type
    Collection
    ...glycosylase, aka T4 Endonuclease V) 165540 pOpen-T7EI T7 Endonuclease I 165551 pOpen-Endonuclease IV E. coli E... E. coli Endonuclease IV 165561 pOpen-Endonuclease VIII E. coli E. coli Endonuclease VIII 165571 pOpen-Exonuclease...pOpen-Exonuclease III E. coli E. coli Exonuclease III 165559 pOpen-T5gene12/13 T5 Exonuclease 165560 pOpen-T4...Collection DNA Ligases DNA Polymerases Restriction Endonucleases Reverse Transcriptases RNA Polymerases Other...below: DNA Ligases DNA Polymerases Restriction Endonucleases Reverse Transcriptases RNA Polymerases Other...phi29 DNA Polymerase Return to top Restriction Endonucleases Addgene ID Plasmid Gene/insert 165504 pOpen-...methyltransferase 165520 pOpen-T4 PNK T4 PNK polynucleotide kinase 165521 pOpen-lambda red operon lambda...

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

    Type
    Blog Post
    ...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...

  6. TALEN Guide

    Type
    Collection
    ...a zinc finger (ZF) nuclease. Zinc fingers recognize triplets and the FokI nuclease operates as a dimer...sites. (B) A TAL effector nuclease (TALEN). Similar in principle to the ZF nuclease, the components of the...the array recognize individual nucleotides. For over ten years, zinc finger (ZF) arrays have been the go-to...number of zinc fingers that recognize various nucleotide triplets. With some trial and error, ZF arrays...years of research has shown that when fused to a nuclease or activation domain, a ZF array is an effective...without their limitations, however. Not every nucleotide triplet has a corresponding zinc finger, and ...Bogdanove at Iowa State University published the nucleotide recognition code of the TAL effectors, which ...

  7. Quick, Versatile Plant Transgenesis with GreenGate Plasmids

    Type
    Blog Post
    ...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. “...

  8. Brain Initiative Collection

    Type
    Collection
    ...10aa-H2B-WPRE3-BGHpA AAV vector for nuclear SYFP2 labeling in neurons. Useful for nuclear isolation and scRNA-seq ...pAAV-hDlx-GiDREADD-dTomato-Fishell-5 Gi-DREADD (P2A) nuclear dTomato expression in forebrain GABA-ergic interneurons...pAAV-hDlx-GqDREADD-dTomato-Fishell-4 Gq-DREADD (P2A) nuclear dTomato expression in forebrain GABA-ergic interneurons...

  9. What's New in CRISPR - December 2019

    Type
    Blog Post
    ...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...

  10. CRISPR 101: Cytosine Transversion Editors

    Type
    Blog Post
    ...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...

  11. CASTing Off for New Shores in Human Genome Editing

    Type
    Blog Post
    ...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...

  12. Build Your CRISPR Vocabulary

    Type
    Blog Post
    ...CRISPR-associated.” The most famous of these proteins is Cas9 nuclease, but there are many other Cas proteins as well...the ability to use an RNA guide to target other nucleic acids—usually DNA—to cut, nick, or bind. DNA CRISPR...genetic record of previously encountered foreign nucleic acids. The unusual sequence of these arrays, with...for studying CRISPR. Spacers: Copies of foreign nucleic acid sequences in the CRISPR array that will become...

  13. What's the Best Way to Elute and Store Your Plasmid DNA?

    Type
    Blog Post
    ... from E. coli is the presence of the protein endonuclease I encoded by the gene endA. Nowadays, the most...gene has been removed/mutated to neutralize the nuclease activity. For strains that still carry this gene...introduced in the purification protocol to remove endonucleases activity. For these reasons choosing the correct...most downstream processing. On the other hand, nuclease free water is a great option because it provides...

  14. CRISPR Challenges: Standardization and Homology Directed Repair

    Type
    Blog Post
    ...on Precision Genome Editing with Programmable Nucleases. Check out #KSgenome on Twitter if you missed ...would open not with an introduction to CRISPR nucleases, but with a session on DNA repair. What stuck ...biophysics can help enhance HDR. CRISPR 101: Ribonucleoprotein (RNP) Delivery The RNP delivery method consistently...Mechanisms of precise genome editing using oligonucleotide donors". Kan, Yinan, Brian Ruis, Taylor Takasugi...

  15. Visualizing Translation at the Single Molecule Level

    Type
    Blog Post
    ...Because it contains a nuclear localization signal, NLS-PCP-GFP then returns to the nucleus to bind cognate ...cell. When cellular mRNAs are digested with ribonucleases, ribosomes protect the segments of mRNA to which...Genome-wide analysis in vivo of translation with nucleotide resolution using ribosome profiling.” Science...

  16. Hot Plasmids - September 2019 - CRISPR, Auxin Inducible Degradation, MerMAID Channelrhodopsins, and Genome Looping

    Type
    Blog Post
    ...widely used genome editing tool. However, Cas9’s endonuclease activity can be limited by the need for a specific...and follow the DNA sequence targeted by the Cas9 nuclease. The Streptococcus pyogenes Cas9 (SpCas9) requires... 2-D linear molecule snaking through our cells’ nuclei--our chromatin is comprised of thousands of loops...provide a 3-D structure for packing genomic DNA into nuclei, but are also thought to play a key role in the...

  17. Improved Plasmid Maps Powered by SnapGene

    Type
    Blog Post
    ...identifies plasmid features that differ by only a few nucleotides, such as the fluorescent proteins EGFP and mEGFP...listing commonly used primers detected in a given nucleotide sequence. The table includes primer names, sequences...determine similarity between a given sequence and nucleotide (BLASTN) or protein (BLASTX) sequence databases...databases. Additionally, align a custom nucleotide sequence against a given sequence using BLAST2. See this...

  18. Multiplex Genome Editing with CRISPR-Cpf1

    Type
    Blog Post
    ...Cpf1 CRISPR Nucleases   Cas9 Cpf1 Components crRNAtracrRNACas9 crRNACpf1 Endonuclease size spCas9...and doesn’t require the expression of other endonucleases, like Csy4, or the inclusion of processing signals...Zhang, F. (2015). Cpf1 Is a Single RNA-Guided Endonuclease of a Class 2 CRISPR-Cas System. Cell,163(3),...

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

    Type
    Blog Post
    ...a CRISPR repeat array near a putative effector nuclease. The Cas13d proteins they identified have little...identified Cas13a-c orthologs, but they do include HEPN nuclease domains characteristic of the Cas13 superfamily...great results with CasRx and AdmCas13d fused to a nuclear localization signal (NLS). In a HEK293 mCherry ...each of four targets, including two mRNAs and two nuclear long non-coding RNAs. One interesting twist to ...

  20. Viral Vectors 101: Types of viruses

    Type
    Blog Post
    ...particles enter the nucleus through nuclear pores and thus do not require breakdown of the nuclear envelope during...during cell division to enter the nucleus as gamma-retroviruses do. Fun fact: Since lentivirus and gamma-retrovirus...
Showing: 81 - 100 of 423 results