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We narrowed to 101 results for: donor

Showing: 21 - 40 of 101 results

  1. Fluorescent Protein Guide: FRET

    Type
    Collection
    ... by exciting the donor and measuring the emission of the acceptor. For a given donor-acceptor pair, the... is non-radiatively transferred from an excited donor fluorophore to an acceptor. Since the transfer ...fluorescent proteins have been developed for use as donor and acceptor pairs in FRET experiments. The efficiency...efficiency strongly depends on the distance between the donor and acceptor molecules and can therefore be used...used to calculate the distances between the donor and acceptor. FRET is often used to study (1) protein-protein...interactions where each protein is separately fused to a donor or acceptor molecule (also referred to as intermolecular...conformational changes within a protein where the donor and acceptor are both fused to the same protein ...

  2. CRISPR 101: Cas9 Nickase Design and Homology Directed Repair

    Type
    Blog Post
    ...using a single-stranded oligonucleotide (ssODN) donor. Using a D10A nickase to insert an EcoRI site in...region using either a top or bottom strand ssODN donor with 40 bp homology arms. The HDR efficiency compared...Fragments) Test both bottom and top strand ssDNA donors (if possible) The text and images in this post...

  3. CRISPR Challenges: Standardization and Homology Directed Repair

    Type
    Blog Post
    ...distance affects editing efficiency. Optimizing Donor DNA for Enhanced CRISPR Genome Editing Chris Richardson...of precise genome editing using oligonucleotide donors". Kan, Yinan, Brian Ruis, Taylor Takasugi, and ...conditional knockout mouse models using long ssDNA donors". Miura, Hiromi, Rolen M. Quadros, Channabasavaiah...

  4. Understanding GPCR Signaling Using TRUPATH

    Type
    Blog Post
    ...Förster Resonance Energy Transfer (FRET), in which a donor fluorophore is used to excite an acceptor fluorophore...that the energy transfer is between a luminescent donor and a fluorescent acceptor. TRUPATH uses an updated...system was developed by constructing 14 luciferase donor Rluc8-Gα chimeras, four acceptor GFP2-Gγ chimeras...

  5. Mobile-CRISPRi: Bringing CRISPRi to Diverse Bacteria

    Type
    Blog Post
    ...cassette flanked by left and right Tn7 end sequences. Donor bacteria with a chromosomal copy of the RP4 transfer...sequence-specific DNA binding protein. Selection removes donor bacteria and recipient bacteria that lack an integrated...bacteria strain of interest by tri-parental mating. Donor bacteria contain a chromosomal copy of the RP4 transfer...genome. Antibiotic plates lacking DAP select against donor bacteria and for recipients with an integrated copy...

  6. Generating Mouse Models Using CRISPR/Cas9

    Type
    Blog Post
    ...double-strand break (DSB) at the target site, and a donor oligonucleotide or plasmid as the repair material...basics To create a mouse model, the gRNA, Cas9, and donor oligonucleotide or plasmid components are brought... al., 2018). Alternatively, the gRNA, Cas9, and donor oligonucleotide can be electroporated into the mouse...carry a frameshift mutation. For knock in with a donor oligonucleotide, we aim to generate 50 to 100 mice...deriving mice carrying the intended mutation. With a donor plasmid, the outcome is less predictable. In our...

  7. Bacterial Expression Systems

    Type
    Collection
    ...fluorescence both from the donor and the acceptor fluorophore by only exciting the donor. In BiFC, proteins of...mRuby2-pBAD Clover (donor) mRuby2 (acceptor) FRET Michael Davidson 18856 pGWF1 ECFP (donor) Venus (acceptor...pFLIP38 ECFP (donor) Citrine (acceptor) FRET Wolf Frommer 65617 pFLIP42 mCerulean (donor) Citrine (acceptor...microscopy. FRET relies on the principle that when a donor fluorophore is excited, it can transfer its energy...Application PI 14030 14031 pCyPet-His pYPet-His CyPet (donor) YPet (acceptor) FRET Patrick Daugherty 18084 54856...54856 pBad-mAmetrine1.1 tdTomato-pBAD mAmetrine1.1 (donor) tdTomato (acceptor) FRET/Dual FRET Robert Campbell...Tsien 54553 54723 mTFP1-pBAD mCitrine-pBAD mTFP1 (donor) mCitrine (acceptor) FRET/Dual FRET Robert Campbell...

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

    Type
    Blog Post
    ...addition of a fluorophore or tag. To utilize HDR, a donor template containing the desired sequence flank by...Nanoblades which carries a gRNA, Cas9, and a HDR donor template DNA can be generated by incubating Nanoblade...Nanoblades facilitated HDR with either a ssDNA or dsDNA donor template. In vivo editing of mouse zygotes and mice...

  9. CRISPR Plasmids - CRISPR Transposases (CAST)

    Type
    Collection
    ...CRISPR-associated transposases (CASTs) for insertion of donor DNA....a guide RNA. The transposase can then integrate donor DNA. Editing by CASTs is not considered "scarless...homology sequences are integrated along with the donor DNA. The two main classes of CASTs are type I and...

  10. CrispyCrunch: High-throughput Design and Analysis of CRISPR+HDR Experiments

    Type
    Blog Post
    ... For each gene, we needed to design guide RNAs, donor templates and sequencing primers for quality control...it ranks guides by CFD score, and it mutates HDR donor templates to avoid re-cutting when needed. It gives...Design gRNAs by inputting the target regions Design donor RNA Design primers Review and order reagents Perform...

  11. INTEGRATE: Bacterial Genome Engineering Using CRISPR-Transposons

    Type
    Blog Post
    ...promoter, and a donor plasmid (pDonor) encodes the mini-transposon. pEffector and pDonor can also be combined...Three transposase proteins (TnsA, TnsB, TnsC). The donor DNA (aka mini-transposon), which contains the DNA...complex with the mini-transposon, excises it from the donor molecule, and inserts it ~50 bp downstream of the...

  12. Plasmids 101: Using Transposons in the Lab

    Type
    Blog Post
    ...double-stranded break, and excises the transposon from the donor DNA. A DNA footprint is left behind. When the transposon-transposase...transposase proteins (TnsA, TnsB, TnsC), and 4) the donor DNA or mini-transposon that contains the DNA cargo... binds the mini-transposon, excises it from the donor molecule, and inserts it ~50 bp downstream of QCascade...

  13. 10 Great Guest Posts We're Thankful For

    Type
    Blog Post
    ...post and the associated publication. Optimizing Donor DNA for Enhanced CRISPR Genome Editing CRISPR has...s lab, describes how you can better design your donor DNA template to achieve more efficient genome editing...

  14. An “elegans” Approach to Better CRISPR/Cas9 Editing Efficiency

    Type
    Blog Post
    ...CRISPR/cas9 and PCR-based homologous recombination donors in cultured Drosophila cells. Nucleic Acids Research...Single-Stranded and Long, Partially Single-Stranded DNA Donors in Caenorhabditis elegans. Genetics 210:781–787...Modifications Improve Potency and Efficacy of DNA Donors for Precision Genome Editing. BioRxiv. https://...354480 Ghanta KS, Mello CC (2020) Melting dsDNA Donor Molecules Greatly Improves Precision Genome Editing...

  15. CRISPR 101: Validating Your Genome Edit

    Type
    Blog Post
    ...Validating knock-in mutations You’ve introduced a donor template along with your gRNA and Cas9 combo and... your edit of interest. Using the same gRNA and donor, you can deliver a silent mutation in a neighboring...that it uses a 3rd sequencing trace file from the donor DNA. This method can precisely estimate editing ...

  16. Fluorescent CRISPR Reporters: SRIRACCHA and GEmCherry2

    Type
    Blog Post
    ...to work and to make the desired genome edit:  A donor plasmid that contains a puromycin resistance gene...by Cas9 and repaired via HDR using the supplied donor plasmid will express GFP. If there’s GFP expression...

  17. Gendered Innovations: Why Does Sex of the Cell Matter?

    Type
    Blog Post
    .... In this case, they found that sex-matching of donor and recipient yielded the best results, but this...forward. It’s just not that easy. All combinations of donor/recipient interaction should be tested before being...

  18. CRISPR in the Clinic

    Type
    Blog Post
    ...Finally, allogenic therapies involve editing healthy donor-derived cells and delivering them to the patient...been modified ex vivo, previously edited healthy donor cells can be delivered immediately, saving precious...

  19. Hot Plasmids - August 2020

    Type
    Blog Post
    ...electroporation and AAV donor infection (CRISPR-READI) enables genome engineering using HDR donors lengths up to...
Showing: 21 - 40 of 101 results