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We narrowed to 402 results for: MUT

Showing: 101 - 150 of 402 results

  1. 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...

  2. 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...

  3. 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...

  4. 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...

  5. CRISPR 101: Any Base Transversion Editors

    Type
    Blog Post
    ...this promising start, the team used rational mutagenesis to produce the adenine transversion editor AYBEv3... edits. Their final tool, ACBE-Q (for an N108Q mutation in the deaminase), produced an average A → C product...and ABEs, they could be useful for saturation mutagenesis, and they provide a strong proof of concept for...empty space. Over several rounds of rational mutagenesis, the team progressed to gGBEv6.3, which was capable...

  6. The PAM Requirement and Expanding CRISPR Beyond SpCas9

    Type
    Blog Post
    ...These off-target sequences may be unintentionally mutated along with the desired target locus. The PAM sequence...positive selection screens in bacteria to identify mutants of S. pyogenes Cas9 that were able to cleave target...PAM binding specificity: SpCas9 variant Mutations (relative to SpCas9) PAM sequence D1135E...human genome. Another variant, xCas9 3.7 has 7 mutations found in the REC2, REC3, and PAM interacting domains...

  7. New CRISPR Tools: Cas7-11 and PASTE

    Type
    Blog Post
    ...patient's specific mutations. With a drag and drop approach, however, the specific mutations wouldn’t matter...which can be caused by thousands of different mutations, all found in the same gene. If we only have small...matter – the mutated copy could be replaced by simply dragging and dropping in a functional copy of the entire...

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

    Type
    Blog Post
    ...codon bias. The idea is that scientists can make mutations throughout a gene of interest based on an organisms...) to name a few. This could affect synonymous mutations which have even been linked to some diseases. ...human genes contain a region where a synonymous mutation can be harmful (Sauna and Kimchi-Sarfaty, 2011... Understanding the contribution of synonymous mutations to human disease. Nat Rev Genet 12:683–691. https...

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

    Type
    Blog Post
    ...engineered by introducing Cas9-mediated frameshift mutations in the gene of interest. The genomic target for...for Cas9 and the introduction of this mutation can be any site early in the coding region of the gene ... experiments may require the introduction of a mutation at a specific site e.g., deleting the catalytic...modification at a defined amino acid to mimic a patient mutation or maybe to disrupt a catalytic residue. Good ...

  10. How to Write a Scientific Review Article

    Type
    Blog Post
    ...studies that reported mutations in that protein (including how frequently a mutation was found and the size...frequently each member of a class of proteins was mutated, as reported in various studies. We made a table...could easily see how frequently each protein was mutated, you could see how big the studies were, and you...

  11. Plasmids 101: E. coli Strains for Protein Expression

    Type
    Blog Post
    ... with sub-optimal codon usage (Table 1) A few mutations are common to all or most expression strains to...levels including:  ompT: Strains harboring this mutation are deficient in outer membrane protease VII, ...methylate DNA. dcm: Similarly, strains with this mutation are unable to methylate cytosine within a particular... of unstable proteins Tuner (DE3)   Contains mutated lac permease whch allows for linear control of ...

  12. Easi-CRISPR: Generating Knock-In and Conditional Mouse Models

    Type
    Blog Post
    ...first generate your desired mutation in mouse ES cells, select for the mutation and inject the ES cells into... models. For simple knockout alleles or point mutations, the 8-10 month process of conventional gene targeting...PAM; if not, you can add a silent PAM or guide mutation. Designing an Easi-CRISPR conditional allele The...

  13. CRISPR 101: Multiplex Expression of gRNAs

    Type
    Blog Post
    ... are available: wt humanized Cas9, D10A nickase mutant (Cas9n), dCas9 (transcriptional repression), and...recombineering, with 96-99% efficiency for point mutations and 66% efficiency for small insertions. Multiplexing...degradation tag (LVA tag) to achieve specific point mutations in E. coli. gRNAs are delivered on a different.... Yin, Linlin, et al. “Multiplex Conditional Mutagenesis Using Transgenic Expression of Cas9 and sgRNAs...

  14. Keeping Up With CRISPR/Cas9

    Type
    Blog Post
    ...that CRISPR's could be used to correct disease mutations, both in mice and in human cells. But that important...genes in one step without producing off-target mutations," Sha was quoted as saying when the news first...blogger Caroline LaManna put it well: "Wow, custom mutations in monkeys already. I know we've discussed it ...

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

    Type
    Blog Post
    ...by blue light (470nm), paCas9-1 induced indel mutations via NHEJ (frequency of 20.5%) and induced modifications...significantly alter the system’s efficiency at generating mutations when activated with light and lowered background...Hemphill et al. demonstrated that the Cas9 K866PCK mutant was indeed inactive prior to irradiation with UV...., 2015   Nitrobenzyl-caged lysine Cas9 K866PCK mutant Genome editing by NHEJ and HDR Jain et al., 2016...

  16. CRISPR Methods for Bacteria: Genome Engineering, CRISPRa, CRISPRi, Base Editing, and More

    Type
    Blog Post
    ...non-edited cells to survive have inactivating mutations in the Cas9 or spacer sequence, and these rare...pneumoniae and can be used to generate multiple mutations simultaneously (Jiang et al. 2013). Genome editing...recombineering, with 96-99% efficiency for point mutations and 66% efficiency for small insertions. Multiplexing...they were effectively able to introduce point mutations in P. aeruginosa, Pseudomonas putida, Pseudomonas...

  17. Choosing Your Perfect Empty Backbone

    Type
    Blog Post
    ...wild-type version of YGOI to rescue a mutated strain or a mutated version of YGOI to test the different...different phenotypes of the specific mutations. For these purposes Addgene has a large collection of yeast expression...

  21. Plasmids 101: Origin of Replication

    Type
    Blog Post
    ...negative regulation and can be manipulated with mutations in the replicon. For example, the pMB1 ori maintains...per cell, while pUC – which differs by only two mutations – will produce as many as 700 copies per cell....for single- and multiple-unnatural amino acid mutagenesis in Escherichia coli. Biochemistry. 2013 Mar 12...

  22. 15 Years of Addgene: The Top 15 Plasmids

    Type
    Blog Post
    ...plasmid of all time, but was found to have a point mutation in the puromycin cassette that makes it less efficient...Cas9, this plasmid expresses Cas9n (a D10A nickase mutant) to nick, rather than cleave DNA. Find pSpCas9n...(BB)-2A-Puro (PX459) V2.0. It contains a point mutation in the puromycin cassette that makes it less efficient...

  24. Back to Bacteria: CRISPR gRNA Multiplexing Using tRNAs

    Type
    Blog Post
    ...vivo and in vitro, as well as to insert targeted mutations through homologous recombination. To further increase...protoplasts, Xie et. al observed slightly higher mutation rates than they’d previously reported in protoplasts..., they found that 50% of plants had biallelic mutations at all five loci. This highly efficient multiplexing...

  26. Addgene's Tips for Plasmid Quality Control

    Type
    Blog Post
    ...plasmid are the backbone, inserted gene/functional mutations, antibiotic resistance, restriction sites used...most sequence assembly errors occur), deletions, mutations, in-frame protein fusions, and multiple cloning...cases, we deal with multiple inserts or internal mutations that require more than two sequencing reactions...

  27. A Look at Addgene's QC Process

    Type
    Blog Post
    ... function of the plasmid. This includes point mutations, frame shifts, missing elements, and nucleotide...plasmid that are less likely to affect function when mutated; elements that are difficult to sequence; approaches...Sequence discrepancies are not always a result of mutations in the plasmid, but can arise due to errors in...

  28. CRISPR 101: Targeting Non-Coding RNAs with CRISPR/Cas9

    Type
    Blog Post
    ...the majority of the lncRNA locus, scientists can mutate the transcription start site, splice sites, exons...function (Hazan and Bester 2021; Zibitt et al. 2021). Mutating splice sites is particularly effective as it can... Transcription start site (TSS) deletion. (B) Mutations of splice sites. (C) Removal of an exon or a large...

  29. Hot Plasmids February 2024

    Type
    Blog Post
    ... generated a series of StayGold mutants and identified a triple mutant, Y187F, R144I and T155Q, which ...Balasubramanian and colleagues showed the single mutation E138D is sufficient to disrupt dimerization while...

  30. Hot Plasmids: Summer 2025

    Type
    Blog Post
    ... during differentiation after they introduced mutations that drive leukemia (Figure 3). Figure...useful for studying the impacts of other cancer mutations and non-genetic diversity in human cancer stem...responses to the acquisition of leukemic driver mutations. Cell Stem Cell, 32(4), 564–580.e6. https://doi.org...

  31. Adenoviral Delivery of CRISPR/Cas9 Aims to Expand Genome Editing to Primary Cells

    Type
    Blog Post
    ...tissues with Cas9 and a gRNA designed to target a mutated gene. In a Scientific Reports paper introducing...-transformed cells yielded rates of targeted mutagenesis similar to those achieved by isogenic AdVs encoding...CRISPR/Cas9 nuclease complexes induces targeted mutagenesis in a diverse array of human cells. Sci Rep 4:...

  32. Fluorescent Proteins 101: Fluorescent Protein Timers

    Type
    Blog Post
    ...knowledge of protein structure and saturation mutagenesis. mK-GO matures from green to red, and the FT ...absorption spectra). Subach et al’s directed mutagenesis also produced the three separate timers shown...3. Verkhusha, Vladislav V., et al. "An enhanced mutant of red fluorescent protein DsRed for double labeling...

  33. PITChing MMEJ as an Alternative Route for Gene Editing

    Type
    Blog Post
    ...this process accounts for a percentage of the mutations seen with TALENs and CRISPR. MMEJ is active during...and Ling Lu. (2016). “Highly efficient CRISPR mutagenesis by microhomology-mediated end joining in Aspergillus...Other Genome Engineering Tools like Sleeping Beauty Mutase Find the Ideal CRISPR Software for Your Experiment...

  34. Hot Plasmids: Fall 2024

    Type
    Blog Post
    ...TBD domains, part of the HB domain, and several mutations to improve solubility and stability (Figure 2)...in natural DNA repair pathways and spontaneous mutations.  Here, the researchers were able to target multiple...abscessus either simultaneously or stepwise, achieving mutation efficiencies of up to about 9% of colonies (102...

  36. Advanced Uses of Cre-lox and Flp-FRT - A Neuroscientist’s View

    Type
    Blog Post
    ...one of the FLP-FRT systems in your experiments. Mutated FRT and loxP sites in Flex Vectors – inversion ...common means to achieve ligand control is to fuse a mutant estrogen receptor (ER) ligand binding domain (LDB...Properties of FLP Recombinase Evolved by Cycling Mutagenesis.” Nature biotechnology 16(7): 657–62. PubMed ...

  37. Antibiotic Resistance: An Old Solution but a New Problem

    Type
    Blog Post
    ...transfer’. Resistance can also arise from random mutations in the genetic sequence that happen to be beneficial... if some didn’t develop, either through random mutation or horizontal gene transfer, means to defend themselves...effective drug targets (ones that are less likely to mutate or change in order for the bacterium to survive...

  38. How to Design Your gRNA for CRISPR Genome Editing

    Type
    Blog Post
    ...fluorescent tag or the introduction of a specific mutation, one generally relies on homology directed repair...experimental conditions used, led to no detectable mutations (Veres et al., 2014). When working with single-cell...Musunuru K (2014) Low Incidence of Off-Target Mutations in Individual CRISPR-Cas9 and TALEN Targeted Human...

  39. When Fidelity Matters: A frank discussion about ligase fidelity

    Type
    Blog Post
    ...Electrochemiluminescent assay for detection of extremely rare mutations based on ligase reaction and bead enrichment."...direct identification of allelic composition of mutated DNA in a mixed population of excess wild‐type DNA...probe for the detection of low-abundance point mutations." Analytical chemistry 86.1 (2013): 900-906. PubMed...

  40. Cancer and the Immune System: Deciphering the Relationship

    Type
    Blog Post
    ...vasculature to feed themselves, use the blood stream to commute to and invade multiple organs, and most importantly..., tumor penetration, inadequate concentration, mutation of cell surface targets by tumor cells, are a ...the most important: How will cancers evolve and mutate based on the immunotherapeutics we use on them?...

  42. CRISPR 101: Cytosine and Adenine Base Editors

    Type
    Blog Post
    ...being able to specify and correct single-base mutations without introducing DSBs at all. Base editing ...properties of the individual Cas9. They also mutagenized the cytidine deaminase portion of the base editor...are smaller and less likely to cause off-target mutations than those derived from naturally occurring cytidine...

  43. Your Top Requested Plasmid in 2016!

    Type
    Blog Post
    ... V2.0 from it's predecessor, which had a point mutation in the puromycin resistance gene that made it ...less effective in certain cell lines. This point mutation has been corrected in V2.0. pSpCas9(BB)-2A-Puro...

  45. What's the Best Way to Elute and Store Your Plasmid DNA?

    Type
    Blog Post
    ...repaired by base excision repair, but can lead to mutations in single stranded DNA undergoing replication....endA negative, meaning the gene has been removed/mutated to neutralize the nuclease activity. For strains...

  46. Running for Rare Disease, Running for FOP, Running for AJ

    Type
    Blog Post
    ...disease typically caused by a single R206H point mutation on the receptor ACVR1. ACVR1 triggers appropriate...regeneration in healthy patients. However, the R206H mutation makes ACVR1 responsive to members of the ubiquitous...

  47. Plasmids 101: Antibiotic Resistance Genes

    Type
    Blog Post
    ...inactivates the gene and acts as a marker for the mutation. When designing these types of experiments it ...not to use the same resistance cassette for the mutation and for plasmid selection. Additionally, scientists...

  48. Plasmids 101: TOPO Cloning

    Type
    Blog Post
    ... Blog Perform Site Directed Mutagenesis by PCR Use REPLACR Mutatgenesis to Easily Generate Insertion ...

  50. Another Pathway into Cells: iTOP

    Type
    Blog Post
    ...toxin, indicating a 70% frequency of biallelic mutations. Importantly, hESCs retained their stemness, indicating... remove the risk of viral integration causing mutations and gene expression changes elsewhere in the genome...
Showing: 101 - 150 of 402 results