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We narrowed to 417 results for: NUC

Showing: 141 - 160 of 417 results
  1. What's New in CRISPR - May 2019

    Type
    Blog Post
    ...enhance HDR repair using a single-stranded oligodeoxynucleotide HDR template covalently attached to the ...the Cas9-gRNA RNP complex via the fused HUH endonuclease PCV. This increases HDR efficiency up to 30-fold...meningitidis, Nme2Cas9, that recognizes a simple dinucleotide PAM (N4CC), lending to high target site density...
  2. CRISPR Protocol for Genomic Deletions in Mammalian Cell Lines [Video]

    Type
    Blog Post
    ...experiments of Kim and colleagues using zinc finger nucleases to harness non-homologous end joining repair (...chromosomal deletions in human cells using zinc finger nucleases. Genome Res. 2010 Jan;20(1):81-89. PubMed. 2. ... interspaced palindromic repeats (CRISPR)/Cas9 nuclease system in mammalian cells. J.Biol.Chem. 2014 Aug...
  3. SunTag and Fluorescent Imaging

    Type
    Blog Post
    ...activation. Multiple nuclear localization signals (NLS) increase protein import into the nucleus. In developing...-molecule imaging deep inside the cell, in the nucleus and cytoplasm. Again, they found that SunTag marked...
  4. CRISPR 101: Cas9 Nickase Design and Homology Directed Repair

    Type
    Blog Post
    ...By mutating one of two Cas9 nuclease domains, researchers created the CRISPR nickase. Nickases create...insert an EcoRI site using a single-stranded oligonucleotide (ssODN) donor. Using a D10A nickase to insert...small insertions or tags (ssODN; IDT Ultramer® Oligonucleotides) Use 100 nt homology arms for large insertions...
  5. Viral Vectors 101: Parts of the AAV Packaging Plasmid

    Type
    Blog Post
    ...its endonuclease activity to generates a ssDNA substrate which is then nicked by its endonuclease domain...of Rep78 and Rep68 contains a DNA-binding and endonuclease domains, all four Rep proteins share a common...
  6. History of CRISPR Cas - A tale of survival and evolution

    Type
    Blog Post
    ...genome. Cas genes, which code for polymerases, nucleases, and helicases, were also disrupted to elucidate...CRISPR loci to generate RNA, which guide a Cas nuclease to the specified exogenous genetic material next...mutation in the PAM sequence, preventing the Cas nucleases from re-identifying it (3). Surprisingly, few ...
  7. Another Pathway into Cells: iTOP

    Type
    Blog Post
    ... Cas9 and a gRNA, which subsequently enter the nucleus and cleave the specified target. Figure adapted...expression is by nature transient, which may lower nuclease toxicity or the potential for off-target effects... Biol. 2000 Jul;10(7):290-5. PubMed. Is VP22 nuclear homing an artifact? Lundberg M, Johansson M. Nat...
  8. Hot Plasmids - January 2023

    Type
    Blog Post
    ...of happen with Cas13. CRISPR-Csm degrades both nuclear and cytoplasmic mRNA with 90-99% knockdown efficiency...CRISPR-Csm presents an attractive alternative for nuclear-RNA targeting, where siRNAs are ineffective and...expression changes of non-target genes and RNA-seq of a nuclear (MALAT1) and cytoplasmic (CKB1) RNA target. Adapted...
  9. 15 Hot Plasmids from 2017

    Type
    Blog Post
    ...editors with editing windows as small as 1-2 nucleotides! To reduce off-target effects associated with...they purified HF-BE3 protein for delivery in ribonucleoprotein particles (RNPs) to both zebrafish embryos...species. This toolkit includes TALEN and CRISPR nucleases, nickases, and gene activation systems as well... the amount of time that Cas9 is active in the nucleus to reduce off-target gene editing. The bacterial...
  10. Hot Plasmids - February 2022

    Type
    Blog Post
    ...of anti-FLAG-FB-GFP binding to FLAG-tag H2B for nuclear localization. (c) Applications of anti-FLAG-FB-...anti-FLAG-FB-GFP labeling various targets with nuclear (top, 4x-FLAG-mCherry-H2B), cytoskeletal (middle, 4xFLAG-mCherry-β-actin... editing, while gene editing can be done via a nuclease-active CasMINI. This small yet powerful system...
  11. CRISPR 101: Non-Homologous End Joining

    Type
    Blog Post
    ...and does not consider the missing or damaged nucleotides that are common to biological sources of DSBs...factors, including polymerases (Pol μ and Pol λ), nucleases (Artemis), and structure-specific end cleaning...the Cas9 DSB (blunt or near-blunt ends without nucleotide damage) such products are rare, probably accounting...
  12. CRISPRainbow and Genome Visualization

    Type
    Blog Post
    ... and its organization within the nucleus, also called the 4D nucleome. Visualizing specific loci has historically...loci, a major benchmark in characterizing the 4D nucleome. CRISPRainbow: Scaling up to 6 colors + white ...
  13. RNA Extraction Without A Kit

    Type
    Blog Post
    ... of biologically active ribonucleic acid from sources enriched in ribonuclease. Biochemistry 18:5294–5299...effect of ammonium sulfate on the activity of ribonuclease A. Biochimica et Biophysica Acta (BBA) - Enzymology...
  14. Viral Vectors 101: An Introduction to AAV

    Type
    Blog Post
    ... As a genome conversion in the transduced cell nucleus, the single-to-double stranded conversion of the...of symmetry. After uncoating in the target cell nucleus, this DNA structure can readily fold into transcriptionally...DNA folds back on itself to become dsDNA in the nucleus and is a good option if you need rapid transgene...
  15. Click Biology: How to Click and Play to Explore Biology

    Type
    Blog Post
    ...cell-friendly conditions. Using the natural amino acids or nucleic acid bases already present throughout living organisms...being accessible simply by dialing up the right nucleic acid sequence (Addgene can help with this!). Besides... under lots of conditions — in the cytosol and nucleus, nearly all cysteines are reduced; in the ER and...
  16. All in a Twist: dsRNA

    Type
    Blog Post
    ...rarely circulates, but upon viral infection, this nucleic acid can build up. The foreign dsRNA activates ...Transposable elements such as long/short interspersed nuclear elements can generate dsRNA by bidirectional transcription...constantly occurring throughout a single cell, so these nucleic acid hybrids are always around. R-loops are particularly...
  17. Arf GTPase Family

    Type
    Collection
    ...centrosomes, the plasma membrane, lipid droplets, the nucleus, and mitochondria. The activity of GTPases is regulated... regulated both upstream (by GEFs, guanine nucleotide exchange factors), and downstream (by GAPs, GTPase...
  18. CRISPR 101: Anti-CRISPR Proteins Switch Off CRISPR-Cas Systems

    Type
    Blog Post
    ...DNA cleavage Example: AcrIIC1 binds to the HNH endonuclease domain of Cas9 and prevents target DNA cleavage...off-target edits of base editors, which convert a nucleotide base pair to another base pair at a specific ...effects and cytotoxicity that result from excessive nuclease activity in tissues (Li et al., 2018). 4. Selection...
  19. Zebrafish Plasmid Collection

    Type
    Collection
    ...site-specific recombination, morpholinos, zinc finger nucleases, TALENS, and CRISPR/Cas9-based gene editing make...Cas9 - Jeffrey Essner Lab CRISPR-Cas/RNA-Guided Nuclease (RGN) expression plasmids - Keith Joung Lab CRISPR...
  20. 3D Printing Meets CRISPR Cas9

    Type
    Blog Post
    ...protein – but were very confused because the HNH nuclease active site wasn’t even close to its reported ... DNA strand, and then activate two different endonuclease active sites. Fortunately, there is never just...displaced non-target DNA strand and shows the HNH nuclease domain swung around in close juxtaposition to ...
Showing: 141 - 160 of 417 results