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We narrowed to 138 results for: THY-2

Showing: 81 - 100 of 138 results

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

    Type
    Blog Post
    ... pCRISPomyces-2 contains Cas9 and a gRNA cassette. The simpler system of pCRISPomyces-2 displays a higher...and a subsequent curing protocol that requires only 2-3 hours incubation. E. coli and T. citrea scarless...four major components including (1) a CRISPR RNA, (2) four proteins forming the QCascade DNA-targeting ...to dCas9 and efficiently introduce cytosine-to-thymine substitutions in target sequences (Banno et al....

  2. Hot Plasmids May 2018 - Optogenetics, Decaffeination, Biosensors, and Fluorescent Protein Tools

    Type
    Blog Post
    ...C-terminus of human codon-optimized  cryptochrome 2 (CRY2) from Arabidopsis thaliana. When exposed to ...material for the bioproduction of therapeutic methylxanthine drugs like theophylline. To facilitate future...enables the degradation of caffeine (1,3,7-trimethylxanthine) and its conversion into xanthine. The majority...constitutive promoter (BBa_J23100) and expresses N-demethylases and other required enzymes, including a glutathione...rescued by the xanthine produced via complete demethylation of caffeine. For example, the Barrick lab was...

  3. Your Top Requested Plasmid in 2015!

    Type
    Blog Post
    ...PMID: 24336571. PubMed Central PMCID: PMC4089965. 2. Sanjana, Neville E., Ophir Shalem, and Feng Zhang...metalloproteinase-9 secretion, and identifying lysine methyltransferase substrates. We’re incredibly happy that lentiCRISPR...

  4. Troubleshooting and Optimizing a Western Blot

    Type
    Blog Post
    ... that all look exactly alike.      Figure 2: Reagent gradient. Here, the lanes are lightly marked...guide. The gel is then removed from the membrane (2) and the membrane is cut so each lane is its own piece...in the bands, you may want to consult Tables 1 and 2 in our Technical Design of a Western Blot post to ...the antibody’s dynamic range. Specifically, the 1:2, 1:1, and No dilution samples all have approximately...conditions are 4 °C overnight or room temperature for 1–2 hours, there is some evidence that antibodies may ...incubation (10–20 minutes) at 70 °C, or a truly lengthy incubation (30–60 minutes) at 37 °C, if you suspect...

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

    Type
    Blog Post
    ...diffusion test or Kirby-Bauer susceptibility test (Figure 2), and requires at least 18-24 hours to obtain results...." London: Wellcome Trust & HM Government (2016). 2. CDC Information Pages on Antibiotics 3. WHO Antimicrobial...flu) Overuse of antibiotics in livestock farming Lengthy courses of treatment with antibiotics These scenarios...

  6. Bacterial Toxin-antitoxin Systems as Molecular Biology Tools

    Type
    Blog Post
    ...mazF gene and a gene for Zeocin resistance (Figure 2). This MazF-ZeoR cassette is flanked by two direct...nucleotide sequences. To counter this, methyltransferases add methyl groups to bases at specific nucleotide...more stable than the methyltransferase. When a plasmid is lost, the methyltransferase is degraded and/or...composed of restriction endonucleases and methyltransferases. Restriction endonucleases cleave DNA at ...the cell grows resulting in the appearance of unmethylated sites in the chromosome. The more stable endonuclease...

  7. Let There Be LITE Plasmids

    Type
    Blog Post
    ...cryptochrome 2 are the chromophores flavin-adenine dinucleotide and methyltetrahydrofolate, which are universally...

  8. K. phaffii: Rising to the Occasion

    Type
    Blog Post
    ...work with Grows extremely fast (doubling time of ~ 2 hours) Amenable to genetic manipulation The first...operates as an obligate aerobe. It’s also a methylotropic yeast, meaning it can utilize methanol as its...peroxisomes are where methanol metabolism occurs in methylotrophic organisms. K. phaffii is an excellent model...

  9. CRISPR in the Clinic

    Type
    Blog Post
    ... Cure for HIV/AIDS. Int J Mol Sci. 2023 Jan 13;24(2):1563. doi: 10.3390/ijms24021563 Resources on Addgene.org.... Finally, allogenic therapies involve editing healthy donor-derived cells and delivering them to the ... have been modified ex vivo, previously edited healthy donor cells can be delivered immediately, saving...clinical trial for SCD in 2022 as well. Hereditary transthyretin amyloidosis (hATTR) hATTR is caused by a variety...

  10. Top 10 Open Science Developments of 2013

    Type
    Blog Post
    ...access to the results of federally-funded research. 2)      In June, the White House honored 13 “Champions... a few minutes to reflect on some of the most noteworthy developments in open science this year. Here ...

  11. Twenty Years of Sharing: Addgene's Viral Vector Service

    Type
    Blog Post
    ...’re delivering their payload to.   Figure 2: Numbers of AAV tools, by category, available from... substrates, light, or engineered ligands (Figure 2). Biosensors, which include calcium and voltage sensors...significant time and removes a variable from the lengthy process of validating and using an AAV in a specific...

  12. Plasmids 101: Stringent Regulation of Replication

    Type
    Blog Post
    ...the chromosome, ending replication.   Figure 2: Prokaryotic chromosomal replication. Adapted from...Microbiology and Molecular Biology Reviews: MMBR, 62(2), 434–464. https://doi.org/10.1128/MMBR.62.2.434-464.1998...contribute to this regulation in different ways: methylating newly synthesized DNA, blocking binding sites...

  13. Site Directed Mutagenesis by PCR

    Type
    Blog Post
    ...Central PMCID: PMC323600. 2. Chester, Nicholas, and Daniel R. Marshak. "Dimethyl sulfoxide-mediated primer...isolated from a methylation competent bacterial strain (e.g. DH5α which is dam+). Methylation deficient bacterial...template. The parent template is removed using a methylation-dependent endonuclease (i.e. DpnI), and bacteria... of the primers. Because you will be using a methylation-dependent enzyme (DpnI) to eliminate the parent...is unique in that it cleaves only DNA that is methylated at the adenosine of the GATC recognition site...

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

    Type
    Blog Post
    ...restrict methylated DNA fhuA2 lacZ::T7 gene1 [lon] ompT gal sulA11 R(mcr-73::miniTn10--TetS)2 [dcm] R... restriction/methylation system. This means the strain can neither restrict nor methylate DNA. dcm: Similarly...Similarly, strains with this mutation are unable to methylate cytosine within a particular sequence. Browse... in the exponential growth phase are alive and healthy, which makes them ideal for protein expression....

  15. Empty Backbones - Choosing Your Perfect Plasmid Backbone

    Type
    Collection
    ...backbone with MS2 loops at tetraloop and stemloop 2 and EF1a-zeo resistance marker pLenti CMV/TO GFP-Zeo... for gene expression Thy1.1 Mammalian pLKO.3 Thy1.1 - Lentiviral shRNA expression Gentamicin...

  16. CRISPR 101: Any Base Transversion Editors

    Type
    Blog Post
    ...of concept for further development. Figure 2: Mechanisms of deaminase-based adenine transversion...hypoxanthine) or between the pyrimidine bases (cytosine, thymine, and uracil). Base transversions are edits that...have an enzyme to remove inosine from DNA: N-methylpurine DNA glycosylase (MPG), also known as alkyladenine...BioRender.com. Base excision editing extends to thymine Since an enzyme that excises inosine can be modified...which excises uracil, could allow it to recognize thymine (T) or cytosine (C). Several different teams succeeded... DNA, creating an abasic site. MPG / AAG N-methylpurine DNA glycosylase / alkyladenine DNA glycosylase...editing by fusing an adenine base editor with an N-methylpurine DNA glycosylase. Nature Biotechnology, 41(8)...

  17. Hot Plasmids: Summer 2024

    Type
    Blog Post
    ... from damage (Abe & Lim, 2024).    Figure 2: A) When proteins (blue) interact with the air-water...editor consisting of dCas9 fused to a DNA methyltransferase — that can drive lasting transcriptional silencing...Histone tail for Autoinhibition Release of Methyltransferase (CHARM), with several advantages over the ...  Rather than overexpressing an exogenous methyltransferase for epigenetic silencing, which can be toxic...tail to recruit and stimulate the endogenous methyltransferase. The strategy is generalizable to a variety...D3L recruits the endogenous, autoinhibited methyltransferase DNMT3A to the genomic location specified by...DNA-binding protein. The ADD domain of the methyltransferase binds to H3K4me0 on D3L, releasing the autoinhibition...

  18. Generating Mouse Models Using CRISPR/Cas9

    Type
    Blog Post
    ...posts: Part 1: Genetically Engineered Mice and Part 2: Breeding and Crossing Mice. My first CRISPR mouse...is less predictable. In our best case, we saw that 2 out of 3 mice carried a 5 kb insertion in the ROSA...:1262–1267 . https://doi.org/10.1038/nbt.3026 Gurumurthy CB, Takahashi G, Wada K, Miura H, Sato M, Ohtsuka...

  19. CRISPR Guide

    Type
    Guide
    ...left free to interact with the target DNA. Figure 2: Overview of the NHEJ repair mechanism Cas9 will only...systems enable researchers to target anywhere from 2–7 genetic loci by cloning multiple gRNAs into a single...Cas9 is included in the gRNA-containing plasmid, or 2-vector systems, in which Cas9 must be delivered separately...your experimental cell population (Figure 8E). In a 2-vector system, you’ll need to either co-infect with...the presence of infectious organisms (like SARS-CoV-2 ) and genetic mutations. Similar to Cas9 and Cas12...Cas enzymes? CasPEDIA is an encyclopedia of Class 2 CRISPR systems with wiki entries describing enzyme...off-target effects by using a single Cas9 nickase and 2 different gRNAs, which bind in close proximity on ...

  20. Transferable Skills Guide: Conflict Resolution

    Type
    Blog Post
    ...become a valuable asset yourself.     Fig 2: A diagram of the Thomas-Kilmann Conflict Model, courtesy...as well. What I really brought to the table was empathy, a willingness to listen, and an ability to remember... are a few more tools that can come in handy: Empathy: If you often find yourself in confrontational ...critical component of conflict resolution. Having empathy does not mean you like everyone you meet. Most ...solid examples of times when you have used your empathy, listening skills, and institutional knowledge ...
Showing: 81 - 100 of 138 results