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We narrowed to 423 results for: NUC

Showing: 141 - 160 of 423 results

  1. 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...

  2. 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 ...

  3. AAV Vector Quality Control: Going the Extra Mile with NGS

    Type
    Blog Post
    ...someone grabs the wrong plasmid? What if bacteria snuck into the prep during the production process? These... Each contig is then BLASTed against the NCBI nucleotide collection database, and it may also be aligned...Next-generation Sequencing. Molecular Therapy - Nucleic Acids 4:e260 . https://doi.org/10.1038/mtna.2015.32...

  4. Google Forums Round Up: First Impressions of NgAgo

    Type
    Blog Post
    ...stage with much-lauded CRISPR-Cas9 is DNA-guided endonuclease NgAgo! We'll discuss how NgAgo is faring with...mammalian cells and one which diminishes the endonuclease activity of the enzyme. In vivo, only nascent...someone came up with a mutant NgAgo (or a novel endonuclease) protein that can form a complex with a 5’-P-ssDNA...

  6. CRISPR 101: Making a Knock-In Cell Line

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    Blog Post
    ...on the size of the knock-in. Single strand oligonucleotide donors (ssODNs) with homology arms as short...need to be used due to synthesis limits of oligonucleotides. These donors traditionally have larger homology...Cas break for HR is to fuse an HR protein to the nuclease. Early HR factors fused to Cas9 have been shown...

  7. CRISPR 101: RNA Editing with Cas13

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    Blog Post
    ...editing with various spacer lengths from 30-84 nucleotides, and designated this system REPAIRv1. Using next-generation...had A-to-I off-target edits around the targeted nucleotide. Introduction of guanine-mismatches in the guide... at Addgene. Gootenberg, Jonathan S., et al. "Nucleic acid detection with CRISPR-Cas13a/C2c2." Science...

  8. New Optimized Genome-wide CRISPRko, CRISPRi, and CRISPRa Libraries

    Type
    Blog Post
    ...commonly result in a complete loss of function while nuclease-deactivated Cas9 (dCas9) can be tethered to a ...which contains multiple point mutations in the nuclease domain. These mutations result in an inactive ...CRISPRa, sgRNAs were targeted between 150-75 nucleotides upstream of the TSS. Similar to the Brunello ...

  9. Tips for Screening with Yeast Two Hybrid Systems

    Type
    Blog Post
    ...modification, protein degradation, access to the nucleus in eukaryotic screens and fusion configuration....cells, fusion proteins must gain access to the nucleus. To circumvent this requirement for transmembrane...system for the pathogenic fungus Candida albicans. Nucleic Acids Res. 2010;38(19):e184. Pubmed PMID: 20719741...

  10. Antibodies 101: Immunoprecipitation

    Type
    Blog Post
    ...target protein interacts with other proteins or nucleic acids. Read on to learn more about this versatile...difficult to release proteins, including those in the nucleus, more stringent buffers may be needed. Since many...also be used to study proteins that interact with nucleic acids. Labs studying epigenetics routinely use ...

  11. Analyzing CRISPR Editing Results with ICE from Synthego

    Type
    Blog Post
    ... the ICE software tool soon. To request other nucleases or other features and functionalities, please ...Guide Target - This is the user-defined 17-23 nucleotide sequence of the DNA-targeting region of the guide...Protospacer Adjacent Motif (PAM) sequence for the nuclease used. Currently, ICE is configured for Cas9 fromStreptococcus...

  12. To Codon Optimize or Not: That is the Question

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    Blog Post
    ... of the tRNA contains the anticodon,  a three nucleotide sequence complementary to the mRNA sequence. ... sequence databases: status for the year 2000. Nucleic Acids Research 28:292–292 . https://doi.org/10.1093...codon usage bias, and its potential applications. Nucl Acids Res 15:1281–1295 . https://doi.org/10.1093...

  13. CRISPR 101: Which Cas9 Do I Choose for My CRISPR Experiment?

    Type
    Blog Post
    ...many cases several exons’ worth of DNA. Any ∼20 nucleotides of DNA can serve as the target for Cas9’s single...specific? Perhaps you need to introduce a single nucleotide modification at a defined amino acid to mimic.... & Liu, D. R. Genome editing with CRISPR–Cas nucleases, base editors, transposases and prime editors....

  14. Plasmids 101: Expressing shRNAs for RNA Interference

    Type
    Blog Post
    ...hairpin shRNA structures that are processed by a ribonuclease called Dicer. In a great simplification of the...promoter Mature siRNAs typically range from 19–22 nucleotides and are 100% complementary to their target sequence...assumptions about miRNA-guided gene silencing. Nucleic Acids Research, 50(2), 617–634. https://doi.org...

  15. Viral Vectors 101: Integrase-Deficient Lentiviral Vectors

    Type
    Blog Post
    ...viral and cellular proteins and trafficked to the nucleus. Since the integrase is non-functional and yet ...If you want your cells to express a protein or nucleic acid for only a limited time, IDLVs are ideal. ... off-target mutagenesis induced by CRISPR-Cas nucleases in human cells. Nature Biotechnology, 31(9), 822...

  16. 3 Tips to Improve HDR Efficiency for CRISPR Editing in Human Cells

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    Blog Post
    ...end-joining (NHEJ) pathway, leading to nonspecific nucleotide insertions, dele­tions or other mutations, referred...repair template, such as a sin­gle-stranded oligodeoxynucleotide (ssODN), which contains the intended sequence...How to Design Your gRNA Delivering CRISPR Ribonucleoproteins Resources on Addgene.org Browse All CRISPR...

  17. CRISPR 101: Validating Your Genome Edit

    Type
    Blog Post
    ...can deliver a silent mutation in a neighboring nucleotide along with your desired edit. This silent mutation...editing by sequence trace decomposition. (2014) Nucleic Acids Research Volume 42, Issue 22, Page e168. ... template-directed CRISPR-Cas9 editing. (2018) Nucleic Acids Research Volume 46, Issue 10, Page e58. https...

  18. CRISPR 101: Off-Target Effects

    Type
    Blog Post
    ...confer specificity of the nuclease; however, many Cas and Cas-like nucleases are promiscuous. In other...effects? The CRISPR-Cas system is an RNA-guided nuclease that either introduces a ssDNA nick, a dsDNA break...

  19. Viral Vectors 101: AAV Variables That Matter

    Type
    Blog Post
    ... cell type, are intermingled in the same brain nucleus: a serotype that works great in a GABA neuron, ...Volumetric Spread in the CNS. Molecular Therapy - Nucleic Acids, 8, 184–197. https://doi.org/10.1016/j.omtn...AAV-Mediated Gene Transfer. Molecular Therapy - Nucleic Acids, 8, 13–25. https://doi.org/10.1016/j.omtn...

  20. Plasmids 101: Using Transposons in the Lab

    Type
    Blog Post
    ...often terminate with a poly(A). ORF2 encodes endonuclease and reverse transcriptase activities.    ...preferred target site for integration is TA dinucleotides and it leaves behind the CAG DNA footprint from... generated by directly injecting DNA into the pronuclei of fertilized eggs, which leads to random incorporation...
Showing: 141 - 160 of 423 results