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We narrowed to 771 results for: INA

Showing: 81 - 90 of 771 results

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

    Type
    Blog Post
    Published
    Dec. 17, 2020, 1:30 p.m.
    ...efficiency. Finally, as shown in Drosophila S2 cells (Böttcher et al., 2014), inactivation of non-homologous...Originally published Jan 27, 2015 and last updated Dec 17, 2020. This post was contributed by Jordan ...heat-shocking to induce the expression of a Cre recombinase contained within the knock-in. Including Cre-...approach – co-CRISPR from Craig Mello’s lab – used inactivation of the unc-22 gene as their selection marker...Andy Fire’s lab used oligo-mediated knock-in of a dominant mutation, known as co-conversion (Arribere et ...by channeling DNA breaks into the homologous recombination repair pathway. Reagents required to perform...), and Victor Ambros and Gary Ruvkun for their seminal work on micro RNAs in C. elegans. The C. elegans...

  2. Plasmids 101: Common Lab E. coli Strains

    Type
    Blog Post
    Published
    Nov. 7, 2014, 2:56 p.m.
    ...Lactose permease inactivated, lactose cannot be taken up by cell mcrA, mcrBC Inactivation of pathway that... is essential for recombination and general DNA repair Reduces plasmid recombination, increases plasmid...stability, reduced recombination relA or relA1 Relaxed phenotype, mutation eliminating stringent factor...them in 1885. E. coli are mainly found in the intestinal tract of animals. There are many different naturally... foreign (methylated) DNA mrr, Δ(mcrC-mrr) Inactivation of pathway that cleaves methylated adenine or...ara disruption of arabinose metabolism pathway Inability to utilize arabinose as a carbon source leuB...leuB β-isopropyl malate dehydrogenase inactivated Requires exogenous leucine source for growth proAB mutation...

  3. CRISPR 101: Cytosine Transversion Editors

    Type
    Blog Post
    Published
    March 25, 2025, 1:15 p.m.
    ...hypothesized that the adenosine deaminase could also perform cytidine (C) deamination, converting the C base to... picture of chromatin than an accessible/inaccessible binary. A word of caution: CGBEs and indels Cytosine...sequences. These first base editors were both deaminases: they removed an —NH2 (amine) group from the ...consisted of Cas9 fused to an engineered adenosine deaminase capable of chemically converting adenosine (A)... started engineering a CBE to be worse at its original job. They removed the uracil glycosylase inhibitor...part of the likely editing mechanism: cytidine deaminase converts C to U, and UNG excises the new U to ...could explain why the edited U base opposite the original G is not primarily converted back to a complementary...

  4. Viral Vectors 101: An Introduction to AAV

    Type
    Blog Post
    Published
    May 30, 2023, 1:15 p.m.
    ...promoters from subretinal injections of neonatal mouse eyes at two different ages." Retinal Degenerative...optogenetics, and chemogenetics experiments. Recombinant AAVs (rAAVs) The primary application of AAV is...meaning that all viral genes except ITRs (inverted terminal repeats – the sequences that flank the deliverable...deliverable DNA) are removed, resulting in recombinant AAVs (rAAVs) with maximum “trunk” space for genetic ...placed in your cell type of interest. Careful combination of promoters, gene switches, and/or serotype ...dividing tissues will lose expression quicker than terminally differentiated, non-dividing cells.   ...size of the ssAAV genome has a mutation in the terminal resolution site (TRS) to form a vector genome ...

  5. Plasmids 101: Dimers and Multimers

    Type
    Blog Post
    Published
    Jan. 10, 2023, 2:15 p.m.
    ...homologous recombination, growing a plasmid in a bacterial strain that is deficient in recombinase protein... sequencing, (which differ from the short-read Illumina MiSeq sequencing that Addgene uses.) However, ...Plasmid multimerization occurs via homologous recombination between two or more copies of the same plasmid...plasmid (Bedbrook & Ausubel, 1976). This recombination event produces a multimeric form of the plasmid with...article.   Long-read sequencing Addgene uses Illumina MiSeq NGS to perform whole plasmid sequencing ...made with BioRender.   Grow plasmids in recombinase-deficient strains Since plasmid multimerization...bi00850a028. PMID: 4878700. Bedbrook JR, Ausubel FM. Recombination between bacterial plasmids leading to the formation...

  6. Prime Editing: Adding Precision and Flexibility to CRISPR Editing

    Type
    Blog Post
    Published
    Jan. 13, 2025, 9:49 p.m.
    ...flexibility to the CRISPR editing world. The approach, originally published in Nature, is a “search-and-replace...HDR is typically less than 10%. In contrast, the original paper introducing prime editing demonstrated ~...into one strand, there’s a mismatch between the original sequence on one strand and the edited sequence...indels by 13-fold (Anzalone et al., 2019). PE3, as originally introduced, relies on the PE2 enzyme. PE4 and...including indels. Therefore, they engineered a dominant-negative mutant of the protein MLH1, a component...and DNA ligases to act first. PE4 and PE5, as originally introduced, rely on the PE2 enzyme. PEmax: Optimizing...(Yan et al., 2024). The team fused La to the C-terminal end of PEmax to produce PE7. PE7 significantly...

  7. Using AAV for Neuronal Tracing

    Type
    Blog Post
    Published
    Aug. 9, 2018, 1:04 p.m.
    ...onwards to other neuronal cells. Synapses at the terminals of axons form connections to dendrites of proximal...outlines neurons from their cell bodies to the terminals of their axons; while retrograde tracing outlines...outlines neurons in the opposite direction, from the terminals of their axons to their cell bodies. Anterograde... are some reports of bi-directionality. For predominately anterograde tracing, biotinylated dextran amines...rhodamine-isothiocyanate (RITC; Thanos et al., 1987) are used. For predominantly retrograde tracing, albumin protein labelled...AAV8, AAV9 Lung AAV4, AAV5, AAV6, AAV9 RPE (Retinal Pigment Epithelium) AAV1, AAV2, AAV4, AAV5, AAV8...conditional mutant mouse brain that expressed Cre recombinase only in pyramidal cells of the hippocampal region...

  8. CRISPR in the Clinic

    Type
    Blog Post
    Published
    Nov. 2, 2023, 1:15 p.m.
    ...post. From Mice to Men When this article was originally written, the main focus was on editing animal... CRISPR’s therapeutic potential in mice with a dominant cataract disorder caused by a single copy of a...edited ex vivo, and then returned to the patient. Finally, allogenic therapies involve editing healthy donor-derived...early-phase clinical trials and have shown promising preliminary results (Dimitri et al). CRISPR engineering of...and USA federal drug approval boards are being finalized, and it is very likely the treatments will be ... Cas family) as part of a cocktail therapy for urinary tract infections. The Cas3 enzymes target several...have experienced hiccups (either clinically or financially), and others we just don’t have the time to talk...

  9. Golden Gate Assembly Upgrades: More Fragments, Faster Assembly, and Higher Fidelity

    Type
    Blog Post
    Published
    Oct. 11, 2018, 12:30 p.m.
    ...pre-determined manner. If the original insert religates into its original vector, it will subsequently ...occasional uncut or cut/re-ligated pGGA destination plasmids. A final validation of the 5-, 12- or 24-fragment...in 5’ four base overhangs after cutting. The destination vector receives the assembly after similar Type...re-digested and "eligible” for ligation into the destination construct. Reactions are often cycled between...acquisition of antibiotic selectable markers by the destination plasmid. Antibiotic selection allowed for high...is dependent on the presence of every module, destination construct and functioning Type IIS restriction... by both advances in the re-engineering of the original BsaI-HF® Type IIS restriction enzyme and the completion...

  10. Optogenetics + CRISPR, Using Light to Control Genome Editing

    Type
    Blog Post
    Published
    Sept. 3, 2020, 12:15 p.m.
    ... variety of combinations to optimize both fusion proteins. The best performing combination was NLS-dCas9...authors fragmented Cas9 into N-terminal (residues 2-713, N713) and C-terminal (residues 714-1368, C714) halves...This blog post was originally written by Caroline LaManna, published Mar 8, 2016. The updated and expanded...two fusion proteins: 1) the genomic anchor - an inactive, dead Cas9 protein (dCas9) fused to CIB1; and ...settled on a different optimal fusion protein combination. The optimized LACE system consisted of: 1) CIBN-dCas9... CIBN-dCas9-CIBN, where CIBN is the N-terminal fragment of CIB1 and it was fused to both the N- and C-...split-Cas9 (FAST) system constitutively expresses the C-terminal fragment of Cas9 while transcription of the N-...
Showing: 81 - 90 of 771 results