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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: 101 - 120 of 423 results

  1. Viral Vectors 101: Viruses as Biological Tools

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    Blog Post
    ...through nuclear localization signals which can traffic their viral particles inside the nucleus, without...access to the host genome requires breakdown of the nuclear envelope, which occurs during mitosis. The most...PMCID: PMC413431. 2. Bukrinsky MI, et al. “Active nuclear import of human immunodeficiency virus type 1 preintegration...

  2. Sharing is Caring: But How to Distribute Open Hardware?

    Type
    Blog Post
    ... the miCube, SQUID (Hongquan, et. al, 2020) or openUC2 (Diederich et. al., 2020) aim to make microscopy.... There are a number of new platforms (such as openUC2, which I am a co-founder of), that aim to support...microscope or open-source modular optical toolbox openUC2 are trying to overcome this hurdle by taking these...He is co-founder of the open-hardware startup “openUC2” and focuses on bringing cutting-edge research ...

  3. What's New in CRISPR - November 2021

    Type
    Blog Post
    ... works best for genome editing  with an active nuclease.  Read the paper in Molecular Cell Find the CasMINI...Programmed genome editing by a miniature CRISPR-Cas12f nuclease Quanjiang Ji’s lab describes an effective miniature...various delivery methods, including plasmid, ribonucleoprotein, and AAV.  Read the paper in Nature Chemical...protect the RNA from degradation by cellular exonucleases and broadly improve editing efficiency. Read...

  4. Plasmids 101: Golden Gate Cloning

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    Blog Post
    ...a few advantages over other cloning methods. Exonuclease-based methods like Gibson assembly require 20...multiple DNA fragments to create TAL effector nucleases. These plasmids were designed to utilize the BsaI...Gate cloning for inserting the appropriate oligonucleotides specifying a gRNA target sequence into a Cas9... Baller JA, Somia NV, Bogdanove AJ, Voytas DF. Nucleic Acids Res. 2011 Jul;39(12):e82. Epub 2011 Apr 14...

  5. CRISPR History and Development for Genome Engineering

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    Collection
    ...genome engineering technologies like zinc finger nucleases (ZFNs) and TALENs. CRISPR: An Adaptive Immune ...Through sequence homology, these crRNAs guide a Cas nuclease to the specified exogenous genetic material, which...the individual crRNAs. (3) The tracrRNA and Cas9 nuclease form a complex with each individual, unique crRNA... alternative PAM sites . Cpf1 , another CRISPR nuclease, recognizes a 5’-TTN PAM, improving targeting ...Cascio VM, Joung JK. 2014. Improving CRISPR-Cas nuclease specificity using truncated guide RNAs. Nat Biotechnol...Shinagawa H, Makino K, Amemura M, Nakata A. 1987. Nucleotide sequence of the iap gene, responsible for alkaline... E. 2012. A programmable dual-RNA-guided DNA endonuclease in adaptive bacterial immunity. Science . 337...

  6. Zhang Lab CRISPR Page

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    Collection
    ... CRISPR-mediated nucleic acid cleavage. We have harnessed the type II CRISPR nuclease system to facilitate...nterspaced S hort P alindromic R epeats) is a microbial nuclease system involved in defense against invading phages...by co-expressing the S. pyogenes Cas9 (SpCas9) nuclease along with the guide RNA. For most applications... have found that adding an additional guanine nucleotide to the 5’ of 20-bp guide can increase targeting... genes. It consists of three components: A nucleolytically inactive Cas9-VP64 fusion A sgRNA incorporating... have found that adding an additional guanine nucleotide to the 5’ of 21-bp guide can increase targeting...GFP-KASH fusion facilitates FACS sorting of cells and nuclei. Return to top AAV - Cas9 mouse CRISPR-Cas9 is ...

  7. CRISPR Methods for Bacteria: Genome Engineering, CRISPRa, CRISPRi, Base Editing, and More

    Type
    Blog Post
    ... Then, pCRISPR is introduced along with an oligonucleotide repair template. Through recombineering, the...VA, Marraffini LA (2014) Exploiting CRISPR-Cas nucleases to produce sequence-specific antimicrobials. Nat...expression using an engineered CRISPR-Cas system. Nucleic Acids Research 41:7429–7437 . https://doi.org/10.1093...antimicrobials using efficiently delivered RNA-guided nucleases. Nat Biotechnol 32:1141–1145 . https://doi.org...tuberculosisusing an efficient CRISPR interference system. Nucleic Acids Res 44:e143–e143 . https://doi.org/10.1093...

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

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    Blog Post
    ...non-coding RNAs (lncRNA). lncRNAs are over 200 nucleotides in length and lack an open reading frame, so ...activation (CRISPRa) are powerful tools. CRISPRi uses a nuclease-dead Cas9 protein (dCas9) and a sgRNA to temporarily...Charpentier E. A Programmable Dual-RNA–Guided DNA Endonuclease in Adaptive Bacterial Immunity. Science. 2012...with the CRISPR/Cas9 system in human cell lines. Nucleic Acids Research. 2014;43(3):e17. doi:10.1093/nar...

  9. Nanoblades: Tiny CRISPR Ninjas for Genome Editing Difficult Cells

    Type
    Blog Post
    ...MLV)-like particles loaded with Cas9-gRNA ribonucleoproteins (RNPs). According to the authors, they named...insertion of a DNA sequence ranging from a single nucleotide change to large insertions like the addition ...intracellular injections into the cytoplasm or pronucleus of the zygote which is both technically challenging...viral-derived Nanoblades loaded with Cas9-sgRNA ribonucleoproteins." Nature communications 10.1 (2019): 45. ...

  10. Plasmids 101: Terminators and PolyA signals

    Type
    Blog Post
    ...post-transcriptional additional of multiple adenine (A) nucleotides to the tail of a messenger RNA transcript. The...protection from degradation, and is integral to the nuclear export and translation processes as well.   ...employ other non-poly(A) transcript-stabilizing and nuclear export elements such as WPRE (5) and CTE (6), and... Gag expression in a context-dependent manner. Nucleic Acids Research. 2000. PubMed PMID:10648781. PubMed...

  11. Twenty Years of Addgene Sharing: CRISPR

    Type
    Blog Post
    ...CRISPR plasmids containing a nuclease into three functional groups: nucleases, modulators, and chimeric ...chimeric effectors. Nucleases generate a classic double-stranded break, while modulators regulate transcription...function, from 2014 to 2023. As you can see, nucleases, the original CRISPR tool, remain the most popular...

  12. PITChing MMEJ as an Alternative Route for Gene Editing

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    Blog Post
    ...such a system is likely to introduce additional nucleotides flanking the insertion. If the DNA ends anneal...MMEJ may also introduce, substitute, or delete nucleotides in addition to the expected deletion, but the...compare to CRISPR sticky-end insertion using the nuclease Cpf1. Since Cpf1 cuts in a staggered pattern, ...this editing platform lies in the diversity of nucleases and their applications. It will be interesting...

  13. Ras Pathway

    Type
    Collection
    ...NF1 Neurofibromin 1 NFE2L2 Nuclear factor, erythroid 2 like 2 NFKB1 Nuclear factor of kappa light polypeptide...Ral guanine nucleotide dissociation stimulator RAPGEF RAPGEF1 RAPGEF2 Rap guanine nucleotide exchange factor...GTPase and is a member of the G protein (guanine nucleotide binding protein) family. GTPases serve as molecular...activating protein 35 ARHGEF2 Rho/Rac guanine nucleotide exchange factor 2 CCND CCND1 CCND2 CCND3 Cyclin... RASGRF1 RASGRF2 Ras protein specific guanine nucleotide releasing factor RASGRP RASGRP1 RASGRP2 RASGRP3...scaffold protein SOS SOS1 SOS2 SOS Ras/Rho guanine nucleotide exchange factor SPRED SPRED1 SPRED2 SPRED3 Sprouty...TSC1 TSC2 Tuberous sclerosis VAV1 Vav guanine nucleotide exchange factor 1 YAP1 Yes associated protein...

  15. Hot Plasmids - August 2020

    Type
    Blog Post
    ...improve nuclear localization of fluorescent proteins, they found that adding a triple nuclear localization...protease allows a transcription factor to enter the nucleus and modulate transcription of the mCitrine gene...includes 95 plasmids consisting of CRISPR/Cas nucleases, base editors, gRNA backbones, and promoters for...

  16. Advanced Uses of Cre-lox and Flp-FRT - A Neuroscientist’s View

    Type
    Blog Post
    ...further improved, by adding a nuclear localization sequence (SV40 Large T nuclear localization sequence) and...sites. FRT sites and loxP sites differ at the nucleotide level but share an overall structure of 13 base...the fusions allowing Cre/FLP translocate to the nucleus where they exert their recombinase activity (Logie...

  17. Tips for CRISPR Gene Editing in Mice

    Type
    Blog Post
    ...target sequence was digested by sgRNA guided Cas9 endonuclease, homology dependent repair (HR, homologous recombination...however, if the microinjection was done as the pronuclei were fusing, you may have several mosaic mice....off-target effects of CRISPR/Cas-derived RNA-guided endonucleases and nickases." Genome research 24.1 (2014): ...Daisuke, et al. "Generation of mutant mice by pronuclear injection of circular plasmid expressing Cas9...

  18. CRISPR 101: Homology Directed Repair

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    Blog Post
    ...can be caused by intracellular factors such as nucleases and reactive oxygen species, or external forces...: The 5’ DNA end of the break is resected by nucleases to create a ssDNA 3’ overhang. This will serve...ssDNA templates (referred to as ssODNs (oligodeoxynucleotides)) are commonly used for smaller modifications...alleles using long ssDNA donors and CRISPR ribonucleoproteins. Quadros RM, et al. Genome Biol. 18(1); (...

  19. Easi-CRISPR: Generating Knock-In and Conditional Mouse Models

    Type
    Blog Post
    ...CRISPR machinery, Quadros et al. recommend ribonucleoprotein (RNP) complexes of Cas9 protein and the separated...alleles using long ssDNA donors and CRISPR ribonucleoproteins.” Genome Biol. 18 (2017) (1):92. DOI: 10.1186...situ germline genome engineering using CRISPR nucleases. Genome Biol. 19 (2018) (1):25. DOI: 10.1186/s13059...Homology Directed Repair Learn about Cas9 Ribonucleoprotein (RNP) Delivery Resources on Addgene.org ...

  20. CRISPR 101: Epigenetics and Editing the Epigenome

    Type
    Blog Post
    ...came epigenome-engineering tools - zinc finger nucleases and TALENs fused to epigenetic modifiers enabled...Christopher Newgard’s Lab has fused LSD1 to the smaller nuclease-dead Staphylococcus aureus Cas9 (Sa-Cas) and deposited... expression and dCas9-based epigenome editing. Nucleic Acids Research 47:e23–e23. https://doi.org/10.1093...CRISPR-Cas9 system for targeted DNA methylation. Nucleic Acids Research 44:5615–5628 . https://doi.org/10.1093...
Showing: 101 - 120 of 423 results