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We narrowed to 399 results for: MUT

Showing: 81 - 100 of 399 results

  1. What's New in CRISPR - November 2021

    Type
    Blog Post
    ...dominant negative mutation in MLH1 (PE4 = PE2+MLH1dn and PE5 = PE3+MLH1dn). Silent mutations added near the...the Cas12f scaffold gRNA and making strategic mutations in the DNA binding cavity of Cas12f. The CasMINI...  and base editing, whereas a different set of mutations (the “V3.1” version) works best for genome editing...boosts the efficiency of prime editing to correct mutations in a variety of target genes and cell types. Their...

  2. Plasmids 101: FLEx Vectors

    Type
    Blog Post
    ...Introduction of a conditional point mutation or functional mutation in vivo: Similar to a conditional rescue...fragment. The availability of both wild-type and mutant loxP sites has allowed scientists to leverage this...wild-type pair (loxP, black arrowheads) and one mutated pair (for example- lox511, white arrowheads). It...conditionally test the function of various point mutations and truncations in the absence of the endogenous...

  3. xCas9: Engineering a CRISPR Variant with PAM Flexibility

    Type
    Blog Post
    ...and select for Cas9 mutants with PAM flexibility. In this system, the mutagenesis plasmid (MP6) permits... Base editing, another method to create point mutations, also has a strict editing window about 15 nt ...NGAG and NGCG and SaCas9 with an NNGRRN PAM by mutagenizing the PAM-interacting domain. However, these alternative...permits quick, large-scale mutation of the gene of interest. They created a selection phage (SP) to carry ...

  4. Polymerase Chain Reaction Overview and Applications

    Type
    Blog Post
    ...colony PCR, or for generating mutated sequences for site-directed mutagenesis.  Quantitative polymerase chain...sequences that are rare, for example, rare alleles or mutations. ddPCR also does not require a reference or standard...interest  Molecular Cloning Site-directed mutagenesis Molecular Diagnostics qPCR Quantify starting... PCR  Plasmids 101: Colony PCR Site Directed Mutagenesis by PCR Resources on Addgene.org Protocol: Polymerase...

  5. Simplify Cloning with in vivo Assembly

    Type
    Blog Post
    ... to the 5’ end of the reverse primer. Mutagenesis - Mutagenesis is performed by designing primers astride...modification, from insertions, deletions, and point mutagenesis to complex assembly of multiple genes. Primer... N-terminus of a protein-coding sequence, or a mutation can be introduced in a gene, while adding a GFP-tag...rapid method for recombination and site-specific mutagenesis by placing homologous ends on DNA using polymerase...

  6. 22 Hot Plasmid Technologies from 2014

    Type
    Blog Post
    ...Updated mini-transposon vector for bacterial mutagenesis or gene targeting Victor de Lorenzo's lab...mini-Tn5 vector that can be used to generate random mutagenesis libraries or to insert heterologous genes, reporters...this vector, called pBAM1, by conducting random mutagenesis in the soil bacterium Pseudomonas putida and ...three dimensional structure of GFP, its brighter mutant S65T-GFP, and the subsequent biochemical characterization...proteins are derived from the Timer protein - a mutant of the fluorescent protein dsRed that changes irreversibly...recombination template vectors, to perform multiple site mutagenesis and create complex fusion or recombination vectors...The group further optimized Chrimson through mutagenesis (K176R) to improve the otherwise slow off-kinetics...

  7. Five Popular Model Organisms

    Type
    Blog Post
    ...different mutations found in leukemia, the Lowe lab was able to identify a set of mutations that made ...Lowe’s lab utilized mice to study how different mutations in leukemia impact different treatment regimens...by a specific gene. For example, several genes mutated in human cancers have a homolog involved in yeast... yeast: A model organism for studying somatic mutations and cancer." Nature Education 1.1 (2008): 183....

  8. Targeting HIV-1 with CRISPR: Shock and Kill or Cut it Out?

    Type
    Blog Post
    ...and CRISPR/Cas9 have also been used to create mutations in genes essential to HIV-1 replication. However...CRISPR therapies through PAM or seed sequence mutations. Multiple gRNAs could be used to combat this problem...LTRs or essential genes. Many of these escape mutations were located near the Cas9 cut site, leading Wang...escape occurs in an animal model. If these escape mutations are frequent, the dCas9-mediated “shock and kill... S, Wainberg MA, Liang C. CRISPR/Cas9-Derived Mutations Both Inhibit HIV-1 Replication and Accelerate ...

  9. Recombinase-based State Machines Enable Order-dependent Logic in vivo

    Type
    Blog Post
    ...which regulatory proteins, signaling molecules, mutations, and environmental conditions drive certain cellular...arrange attB-attP pairs on a register to record all permuted substrings of a set of chemicals. Specific examples...arrangements that should be able to record all permuted substrings of up to seven chemical inputs are ...a population to correctly record each possible permuted substring of the inputs, and that five hour inductions...percent of cells to correctly record each possible permuted substring of inputs (see Figure S9 of our paper...

  10. PiggyBac-ing Through the Genome Editing Field

    Type
    Blog Post
    ...completely seamless manner, leaving no sequences or mutations behind. Furthermore, piggyBac™ offers a large ...is constitutively-active or a dominant-negative mutant. Reducing the copy number can also mitigate the...excision-only piggyBac™ transposase (PBx), which contains mutations making it competent for excision, but defective...primarily two other ways to create specific point mutations by gene editing. First, instead of using piggyBac...piggyBac™ transposase or hyperactive transposase mutants [7], it is generally not as efficient as Cre recombinase...

  11. A History of Genome Engineering in Popular Culture

    Type
    Blog Post
    ...genome and are inefficient at introducing point mutations. In 2016, the researchers at the Broad Institute... envisioned a new strategy to introduce point mutations in the genome. They fused a catalytically impaired...a tRNA deaminase enzyme, TadA) to induce a A→G mutation at the target genomic loci. Together, CBE and ...the prime editor can install all possible point mutations, delete short stretches of DNA and even insert...doi.org/10.1038/nprot.2013.143 Seligman LM (2002) Mutations altering the cleavage specificity of a homing ...

  12. Fluorescent Proteins 101: Photoactivatable Fluorescent Proteins

    Type
    Blog Post
    ...from Jellyfish (Aequorea victoria) GFP (wtGFP) by mutating Threonine 203 to Histidine (T203H) (1). wtGFP ... when excited with 488nm blue light. The T203H mutation in PA-GFP generates a chromophore population that...PA-FPs by performing several rounds of random mutagenesis screening for enhanced RFP variants. PA-mCherry...negative” photoswitching. In 2008, scientists used mutagenesis screens to find a version of Dronpa that doesn't... is a monomeric, reversible PA-FP derived from mutating Dronpa (T59M / V60A / N94I / P141L / G155S / V157G...

  13. 28 Hot Plasmid Technologies from 2015

    Type
    Blog Post
    ...carried out five rounds of random mutagenesis on a library of Arch mutants screening for improved brightness...brightness. The brightest mutants were then further mutagenized and screened for speed and voltage sensitivity...retention insensitivity to other ions In BRET, A mutant Renilla luciferase, catalyzes the oxidation of ...transferred to the acceptor fluorophore, a circularly permuted Venus. In the case of pHlash, the signal from ... SalI sites must be removed by site-directed mutagenesis in order to use the module as a building block...superior to Arch). Both QuasAr1 and QuasAr2 possess mutations for improved voltage sensitivity and speed, and...

  14. How to Deposit Your Plasmids with Addgene

    Type
    Blog Post
    ...Relevant mutations/deletions" box to describe any mutations within your insert using standard mutation notation...fusion proteins contained in the plasmid, any mutations, and the intended purpose of the plasmid. Once...functional consequences of that change. For example, "Mutated Aspartate 123 to Lysine (D123K), lowers biosensor...

  15. 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...SpCas9 nickases as examples for this post. The D10A mutation inactivates the RuvC domain, so this nickase cleaves... only the target strand. Conversely, the H840A mutation in the HNH domain creates a non-target strand-...there are no available guides close to the desired mutation site. Quick tips for nickase design Ready to ...

  16. CRISPR 101: Non-Homologous End Joining

    Type
    Blog Post
    ...processing of DNA ends tends to be the point where mutations are introduced. Repair of Cas9-induced breaks ...about a two-thirds chance of causing a frameshift mutation. Of some importance, the deletion can be less ...double nickase” strategy vastly reduces breaks and mutations at off-target sites. Christina Mork contributed...Persistence of a Chromosome Break Diminishes Its Mutagenic Potential. PLoS Genet 5:e1000683 . https://doi.org...

  17. Magnetic Control of Proteins: More than a Dream

    Type
    Blog Post
    ...more rounds of mutagenesis, they eventually found a version of AsLOV2 with five mutations (C450P, L496V... AsLOV2. After several rounds of semi-random mutagenesis and screening, the EGFP and mScarlet variants... and ΔF/F of MagLOV, a variant of AsLOV2 with mutations C450P, L496V, Q513K, G528K, D540M, in response...

  18. CRISPR 101: Mammalian Expression Systems and Delivery Methods

    Type
    Blog Post
    ...expression? Are you trying to create a single point mutation? Do you want to create a fusion with a reporter...generating knockouts or knock-ins, or introducing mutations and tags, while a catalytically dead dCas9 can... Base editors can help you make precise point mutations near the gRNA target site. Choose an appropriate...repair (HDR) template to create knock-ins, point mutations, or to add a tag, you will also need to deliver...

  19. Plasmids 101: Choosing an Antibiotic Resistance Gene

    Type
    Blog Post
    ...cleavage in human cells. Mutagenesis, 20(2), 111–114. https://doi.org/10.1093/mutage/gei016I Resources on ...when not 100% inhibited, meaning it will induce mutations in the host/plasmid DNA during selection if it...good choice for experiments in which off-target mutations are undesirable. Pros: Works across many species...

  20. 15 Hot Plasmids from 2017

    Type
    Blog Post
    ... used a combination of deliberate and random mutagenesis to isolate the best RFP. They targeted known ...researcher’s locus of interest. NHEJ- or HDR-induced mutations in ATP1A1 can render cells resistant to the small...improves editing efficiency. Importantly, the point mutations created by Agudelo et al. prevent ouabain binding...properties of the individual Cas9. Kim et al. also mutagenized the cytidine deaminase portion of the base editor...FLAG-tagged TZAP constructs (wild-type and truncation mutants), gRNAs used for TZAP gene editing, and a Hi6xs-MBP...
Showing: 81 - 100 of 399 results