Skip to main content
Addgene

Enter a sequence to perform a BLAST-based search of Addgene plasmid sequence databases.

In Category

We narrowed to 974 results for: lat

Showing: 241 - 280 of 974 results

  1. Minigenomes - a Safe Way to Study Dangerous Viruses Like the Ebola Virus

    Type
    Blog Post
    ...terminate transcription and polyadenylate the mRNA. These mRNAs are translated by cellular host machinery...gene start) and a transcription termination/polyadenylation signal sequence (gene end). A minigenome is...minigenome are transcribed (and the viral mRNAs are translated) by cellular machinery. NP encapsidates the negative...then recognizes the encapsidated minigenome as a template for replication and transcription, as it would... necessary control elements for each would be relatively large. Cloning the viral genes in this plasmid...-minigenome). The anti-minigenome is used as a template by the viral polymerase to synthesize additional...addition, the polymerase will use the minigenome as a template to transcribe the reporter gene (e.g. firefly ...

  2. Tips from the Repository Trenches: Using Barcodes to Track Samples

    Type
    Blog Post
    ...location which can all be extrapolated from the 2D barcode and LIMS: Barcoded Tube Plate Freezer Rack Freezer... location within a plate that can hold up to 96 tubes (see image below). Each plate has numbers and letters... Each sample that is transformed is plated onto a petri dish plate that is also barcoded. The barcodes... to give coordinates, from A1 through H12. Each plate is also barcoded with a 10 digit barcode and then...rack that contains five levels that can hold five plates on each level. This rack is located in one of five...sample type. It is critical for us to know which plate, rack, and freezer each sample is located in. Otherwise...identifies the contents of each individual tube.  If a plate of samples is ever dropped, we can simply scan the...

  3. Using Video to Share Your Science: We Share Science

    Type
    Blog Post
    ...not hosted on the platform, they can be hosted on YouTube, Vimeo, and other platforms (such as university...economics, or maybe even biology or international relations.  Many of the challenges in science today are ...appealing to many audiences. While all of these platforms were developing, Google acquired Youtube and has...quickly. Taking what I loved from each of these platforms (such as social networking, visual appeal, instant... across multiple other sites), I created a new platform for sharing video abstracts about research: www.WeShareScience.org...www.WeShareScience.org. WeShareScience is a free platform for sharing short, 3 to 5 minute videos about ...university websites). WeShareScience is a social platform that provides links to the videos, keeping the copyright...

  4. Hassle-free 96-well Format Epitope Tagging Using Cas9 Ribonucleoprotein

    Type
    Blog Post
    ...to the template strand. Therefore, I suggest that picking a crRNA complementary to the template strand...bound by a protein of interest are pulled-down and later coupled to high-throughput sequencing to identify...optimized to achieve high knock-in efficiency. To calculate the tagging efficiency in the transfected cells...tagging experiments by assembling csRNPs in a 96-well plate format and tested parallel tag knock-in for 96 different...single go (Figure 3). We were able to tag >30 genes/plate in the first shot with typical knock-in efficiencies...knock-in experiment were pre-assembled in a 96-well plate. Glioma stem cells were transfected in 96-well format...device; 5 days after the transfection, the entire plate was scanned for V5 tagging using an Operetta imaging...

  5. K. phaffii: Rising to the Occasion

    Type
    Blog Post
    ...for post-translational modification of recombinant protein products, specifically glycosylation. The native...eukaryotic model organism – its genome is easy to manipulate, it’s affordable, and it grows fast to boot. ...doubling time of ~ 2 hours) Amenable to genetic manipulation The first eukaryote (S. cerevisiae) to have...glucose or glycerol). K. phaffii is only distantly related to S. pombe and S. cerevisiae; evolutionarily it...native glycosylation machinery of K. phaffii lacks certain terminal mannose glycopeptides (which most ...in K. phaffii since it produces ‘human-like’ glycosylated proteins. Peroxisome studies Peroxisomes are...biogenesis, degradation, and proliferation. Genome manipulation K. phaffii’s genome has been sequenced for some...

  6. Addgene-Seeding Labs plasmid grant round 3: microbial drug resistance and biofilms, antidiabetic plant components, visualizing brain injury - and a glowing robot fish

    Type
    Blog Post
    ...microscopy methods will be used to stimulate and manipulate neural populations in order to get new insights ...current research, Sheen aims to identify new genes related to PZA-resistance by using different approaches...understanding of PZA action and could potentially stimulate the development of useful diagnostic methods and...collection of pathogenic Klebsiella and Pseudomonas isolates causing urinary tract infections. This project...to create gene deletions of important unknown regulatory regions in Pseudomonas and Klebsiella spp. The... several diseases. Her goal is to identify and isolate pure plant compounds with antidiabetic properties...Recombinant human proteins that play a role in insulin regulation and diabetes will be expressed from plasmids ...

  7. Addgene Welcomes John Doench to the Advisory Board

    Type
    Blog Post
    ...of how miRNAs regulate their targets and what confers specificity in the miRNA regulatory process. This... the then RNAi Platform, which has since evolved into the Genetic Perturbation Platform and encompasses...Associate Director of the Genetic Perturbation Platform at the Broad Institute and has worked with many... up on a Saturday at like 7 am to go check your plates and you’re excited to do so, you know that that...you.” After recognizing his love for science and later realizing that he wanted to continue doing research...The model that the Broad was building that had Platforms with a capital ‘P’ focused on technology ... has...enjoys working on technology development under the Platform model at the Broad Institute because they are ...

  8. Hot Plasmids: Summer 2024

    Type
    Blog Post
    ...the recombinase handles the installation of the donor sequence. The latest work evolves and engineers ...D3L, releasing the autoinhibition and stimulating the methylation of nearby CpG sites on the target DNA...problematic for low-abundance targets or natively isolated complexes. To address these limitations, the Funabiki...Funabiki Lab developed a technique called Magnetic Isolation and Concentration (MagIC)-cryo-EM that combines...streptavidin-coated magnetic beads through an assembly of biotinylated SpyTag-SpyCatcher linkers and spacer proteins...in sample loss. Ci Ji Lim’s lab found that adding Late Embryogenesis Abundant (LEA) proteins, small proteins...particle orientation distribution and Fourier-shell correlation curve (D), and the reconstructed 3D map with ...

  9. Antibodies 101: Beyond Surface Labeling

    Type
    Blog Post
    ...receptors, or post-translational modifications on those proteins, like glycosylation patterns. Surface ...only be interested in one of the various cell populations; for example, CD8+ T (“T killer”) cells. Typically... simply “dump” them by gating on the negative population.     Figure 2: Dump gating labels all...allowing for a flow analysis to be shifted to a later day. Note that intracellular labeling must be performed...method, differences in proliferation between cell populations can be visualized, achieved through, for example...heterogenicity of proliferative behavior in a cell population can be seen as a stepwise decrease in signal ...easily gated on during the analysis. Conclusion Congratulations! You made it through this blog post and hopefully...

  10. SARS-CoV-2/COVID-19 Detection Methods Based on CRISPR/Cas

    Type
    Blog Post
    ...detected using lateral flow strips. Since the probes are tagged with streptavidin, the lateral flow strip ...concept to lateral flow strips that contain streptavidin which can bind to the biotinylated targets. They...earlier epicenters of the outbreak ultimately flattening the curve, it has become evident that widespread...fluorescence and this approach can be incorporated into a lateral-flow readout slip that allows rapid detection.  ...substrate, gets activated and performs non-specific collateral cleavage. This cleavage is coupled to fluorescence.... In this technique, however, instead of using lateral flow strips to detect the cleavage of fluorescently...hours Equipment needed: PCR machines/Heat blocks, Lateral flow strips, Agarose gel equipment, Fluorescent...

  11. Cas13d: Small RNA-targeting CRISPR enzymes for transcriptome engineering

    Type
    Blog Post
    ...cleavage promotes collateral cleavage of other RNAs. As with other Cas13s, this collateral cleavage does ...These small, but mighty type VI-D enzymes are the latest tools in the transcriptome engineering toolbox....identified other advantages, including Cas13d-specific regulatory proteins and high targeting efficiency, both ...accessory protein RspWYL1 increases both targeted and collateral RNA degradation by RspCas13d. RspWYL1 also increased...WYL domain-containing proteins may be broader regulators of Cas13d activity. This property makes WYL proteins...counterpart to anti-CRISPR proteins that negatively modulate the activity of Cas enzymes, some of which are...Konermann et al. compared CasRx to multiple RNA regulating methods: small hairpin RNA interference, dCas9...

  12. Announcing the Winners of the 2021 Michael Davidson and Roger Tsien Commemorative Conference Awards

    Type
    Blog Post
    ...Roger Tsien Commemorative Conference Awards. Congratulations to the five winners! Abhi Aggarwal Brandán... a conference of their choice while honoring the late Michael Davidson and Roger Tsien who have played...thiol-containing acceptors. These hydropersulfides modulate protein activity and have important roles during...opioid use disorder. Her goal is to identify cell populations in the brain that contribute to the development...functional contribution to a variety of behaviors related to addiction and pain. Once these are completed...neurons using GCaMP7, measuring calcium signaling related fluorescence during morphine administration and...resulted in mechanical allodynia. With the dual regulation, she could then use pulses of blue light to activate...

  13. Supporting Rare Disease Research with the 2020 BeHEARD Awards

    Type
    Blog Post
    ...plasmid-based tools for rare disease research. Congratulations to the winners! Below, we highlight their research...patient-mutation bearing knock-in mouse model for CTD. Congratulations the Association for Creatine Deficiencies! ...identify therapeutic targets in the future. Congratulations Mark Mellett! Siddharth Prakash: X chromosome...50-100 times higher than women in the general population,” says Dr. Siddharth Prakash, an associate professor...The lab hypothesized that this is abnormally deregulated in TS and can lead to cardiac developmental defects...cause heart problems in TS,” says Prakash. Congratulations Siddharth Prakash! Terry Hébert: How GNB1 mutations...GPCRs) detect molecules outside of the cell and translate those signals into cellular responses. For example...

  14. Troubleshooting Your Plasmid Cloning Experiment

    Type
    Blog Post
    ...of cells. The antibiotic in your plate might not work. Check the plates! Streak on them an E. coli  strain...help you at this stage and that can be used to simulate in silico cloning. This will definitely help in...enzymes that you are using are not blocked by methylation like XbaI, ClaI, etc. If you are doing multiple...Design primers that overlap at least 24 bp with the template and if the sequence has high GC or AT content,...if you are having difficulties in amplifying a template, particularly from a large genome, you can recheck...have no colonies, check that the antibiotic in the plate correspond to the antibiotic resistance marker present...growing fine, it is probably time to prepare new plates with a fresh stock of antibiotic. Many small colonies...

  15. Advice on Career Paths and the Green Card Process for International Researchers and Entrepreneurs

    Type
    Blog Post
    ... as law, medical/technical writing, clinical, regulatory, product development, business development, consulting...foreign workers in specialty occupations. US regulations define a "specialty occupation" as requiring ...paths are not the only place to develop “industry related skills”. Many career paths in academia as well ...look broadly at opportunities to develop skills related to their areas of interest. Additionally, since...institutions of higher education nonprofit entities related to or affiliated with institutions of higher education...applicant works at institutions of higher education, a related or affiliated nonprofit entity, or nonprofit research...awards, media reports, and membership in groups related to the foreign national’s field.” (Matter of Dhanasar...

  16. Lab Automation at Addgene

    Type
    Blog Post
    ...wells plates onto agar q-trays where the samples must be spread much farther apart before plating. With..., we can prep two 96 well plates in just under 3 hours or one 96 well plate in about an hour. Automating...Beckman Coulter’s Biomek FXP. We had a previous relationship with Beckman Coulter and that certainly played...too much and dying.  Also, we added a couple of platforms that fit 48 well q- trays allowing us to do up...96 tips for easy sample transfer between 96 well plates. The second pod can pick up 8 tips and position... robot is called the Qiagen BIOROBOT Universal Platform and it suits our plasmid prepping needs perfectly...on a larger scale. Using the BIOROBOT Universal Platform is ideal because it eliminates the potential of...

  17. Early Career Researcher Toolbox: Finding Relevant Papers

    Type
    Blog Post
    ... real challenge is finding relevant papers. The latest installment of the Early Career Researcher Toolbox...highlight four tools for finding journal articles related to your actual interests while also staying on ...around since 1996, but is getting an update! The latest version can be found here. The new PubMed will ...these types of accounts can help you find papers related to a particular topic or that were published by...tweet about papers too. You can follow journals related to your field of research or larger journals that...can bookmark a tweet in order to easily access it later. Pro tip Checking the number of retweets, likes,...who published the most papers with your keyword populated at the top of this filter. A unique feature of...

  18. Adapting Toehold Switches to Detect Zika Virus

    Type
    Blog Post
    ...of an outbreak of the virus in Brazil that was correlated with greater rates of infection and rare, but...Zika virus is expected to expand into heavily populated regions of South, Central, and North America. .... The first was a new class of programmable riboregulators, called toehold switches, which allow for the... 2014). The second was an in vitro paper-based platform that allowed gene circuits to be deployed out ... had been working on advancing the paper-based platform toward a more field-ready format when the Zika...our manuscript to the journal Cell just six weeks later. The Zika publication timeline I think we can speak...we say the timeline to develop our paper-based platform into clinically relevant diagnostics for the Zika...

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

    Type
    Blog Post
    ...in 16 wells of a 96-well plate, allowing 6 conditions to be screened per plate. The results indicated that...enzymes that seal breaks in DNA by joining 5 ́-phosphorylated DNA termini to 3 ́-OH DNA termini (1-4). In...interrogation of multiple potential SNP sites (15). The related LCR technique uses two sets of probes to achieve...the reaction. This “peeling back” of the 5 ́-phosphorylated base can be observed in the crystal structures... pools were prepared consisting of one target (template) strand and four upstream probes and four down-stream...strand covering all 16 possible NN pairs in the template. The probes were designed such that each possible...to screen additional ligases, conditions, and formulations and has led to the development of the new HiFi...

  20. Sequencing Options for CRISPR Genotyping

    Type
    Blog Post
    ...genotyping, is critical to demonstrating causal relationships between genotype and assayed phenotype. Verifying...and T7E1 have been widely adopted due to their relative simplicity and low cost. The problem with these...editing events that occur in less that 5% of the population (Fu et al. 2013, Vouillot et al. 2015). Meanwhile...consuming and cannot be applied to heterogenous populations (Bell et al. 2014). Further, Sanger sequencing...Low-throughput, inappropriate for heterogenous populations, low sensitivity Targeted Amplicon Sequencing...list of putative off-target sites that they can later sequence for mutations. One method a researcher ... detection rates mean the investigator can be relatively certain that their samples don’t have off-target...

  21. 22 Hot Plasmid Technologies from 2014

    Type
    Blog Post
    ...phosphoproteins Protein phosphorylation is one of the most abundant forms of post-translational modifications in...questions surrounding serine phosphorylation by adding this phosphorylated amino acid to the genetic code...participates in y. Depletion of your protein may upregulate an alternative pathway or induce other compensatory...cloning have been utilized to create recombination template vectors, to perform multiple site mutagenesis ...where signaling dynamics are influenced by the relatively long life-cycle of a protein from expression ...plasmids required to make singly or multiply phosphorylated proteins (B40 OTS and pCRT7-GFP). To demonstrate... either one or two phosphoserine residues cotranslationally inserted in their canonical positions (SP218...

  22. Custom CRISPR Screens & the Green Listed Software

    Type
    Blog Post
    ...investigator generates a cell population where all genes are knocked out at the population level, but each individual...be targeted. Cells failing to upregulate the protein following stimulation would then be separated by sorting...knocked out. In this genetically heterogeneous population, cells behaving differently (e.g. different growth... response to a cytokine known to cause the up-regulation of an important surface protein (e.g. PD-L1 on...were kindly donated by six different research constellations: Doench/Root (2), Zhang (3), Wu (4), Yusa (...The different labs use different algorithms to calculate which would be the best gRNAs. Interestingly, ...your own favorite Reference Library, for example related to strategies beyond SpCas9, or species not currently...

  23. Rett Syndrome: A History of Research and Therapeutic Outlooks

    Type
    Blog Post
    ...medical journal. Decades later and after years of painstaking work trying to isolate the causative gene for...role of the MECP2 gene, and highlights some of the latest breakthrough therapeutic approaches. What is Rett...et al., 1999). We now know that MECP2 binds to methylated CpG dinucleotides and CAC trinucleotides during...& Bird, 2019) and is a global transcriptional regulator of thousands of genes. Studies have suggested ...with the minor groove of AT-rich DNA sequences to regulate transcription (Baker et al., 2013). Mutations ...in animal models, Rett syndrome is a strong translational candidate for gene replacement therapy. In 2014... expand as Rett scientists deposit their MECP2-related plasmid tools with Addgene and share additional...

  24. Chemogenetics vs. Optogenetics: Which Method Should I Choose?

    Type
    Blog Post
    ...optogenetics differ are: Timing Targeted manipulation Controlling stimulation Invasiveness  Choosing the best...Controlling Stimulation Some studies rely on delivering reversible or varied amounts of stimulation to receptors...where drawn out effects are required. Targeted manipulation Both optogenetic and chemogenetic tools can ...as the light source can be easily and quickly manipulated or turned off. In chemogenetics, while different...injected, this is not as precise and the offset of stimulation is gradual. Invasiveness Both optogenetic and...intracranial implant Noninvasive Controlling Stimulation ExogenousEasily controlled EndogenousNot easily...blog posts Learn more about precise targeted manipulation Resources on Addgene.org Browse all viral ...

  25. A New Optogenetic Tool Based on AraC Controls Gene Expression with Blue Light

    Type
    Blog Post
    ...arabinose making it difficult to regulate quickly. Arabinose also circulates within growth media and can’t...created a bacterial lawn on an agar plate, put a mask over the plate containing the pattern of the image...image, and shined blue light over the plate. Areas on the plate that are blocked by the mask will not produce...May 13, 2021 The original AraC transcriptional regulator Their system is based on the AraC protein that...transcription.However, this system lacks spatio-temporal regulation, and is difficult to reverse, making it unsuitable... show that you have less heterogeneity in the population and you can do all of these more spatially confined...

  26. CRISPR 101: Validating Your Genome Edit

    Type
    Blog Post
    ...worthwhile to isolate individual clones from that population. You can additionally calculate the minimum...used as a preliminary validation on your bulk population. This way you can ensure you aren’t looking for...performed yielding a graph of edits in the input population or clone.   How to perform a TIDE First, ... sequence the PCR product of both an unedited population and your Cas9-targeted cells. Simply upload both...Validating knock-in mutations You’ve introduced a donor template along with your gRNA and Cas9 combo and are hoping...precisely estimate editing frequency in a bulk population and also identify edits in individual clones....reads from your edited sample to an untreated population. Software such as CRISPResso can help with the...

  27. Fluorescent Protein Travel Awards - Protein Variants, a Serotonin Sensor, and an Artificial Leaf Replica System

    Type
    Blog Post
    ...is induced prior to inoculation with IPTG. In the absence of IPTG after inoculation, no more fluorescent...also be used to track how one population responds to a second population. Last, Bernach will simultaneously...The late Michael Davidson and Roger Tsien played enormous roles in fluorescent protein development by...serotonin is a well-known but poorly studied neuromodulator, and evidence suggests that it is incredibly...were able to use the serotonin sensor in sleep-related studies and in assays for measuring human serotonin...nucleotide polymorphisms present in the human population can affect drug and dose response. Selective ...validated, Bernach plans to use it study bacterial population dynamics with the TIMER growth rate bioreporter...

  28. Addgene Depositors Get More Citations

    Type
    Blog Post
    ...demand for the related plasmid. Under such circumstances, we would observe this same correlation, but it wouldn...here? Do papers associated with Addgene deposits accumulate more citations than those without Addgene deposits...aggregated this much, you can see the 2013 ‘bump’ related to the CRISPR/Cas papers and Addgene deposits, ...Addgene, in contrast to other reasons for the correlation. However, determining this exactly is difficult... of publication. Table 1 provides an example calculation of the weights. Table 1: Journal Weighting Example... remaining difference caused by depositing the related plasmids with Addgene? Probably not all of it. ...still be controlled for. As an example, plasmid-related areas of science could be growing more quickly ...

  29. Cancer, Inflammation and Immunity - Harnessing the Body’s Defenses to Fight Cancer

    Type
    Blog Post
    ...human biopsy analysis platform focused on analysing immune measures and correlating them with tumor mutations... immunosuppressive measures, for example the upregulation of the immunosuppressive cell surface receptor...concluded “Aspirin promotes autophagy, inhibits platelet aggregation, and potentiates the effect of PD-...Cancer and the Immune System: Deciphering the Relationship” by our guest blogger Subhadra Jayaraman, a ...Chimeric Antigen Receptors) CAR T cells and PD-1 modulators. An exciting development in immunotherapy is ...mutations and clinical responses. This platform has the potential to become an open-source pipeline and ...spatiotemporal precision. This is a great tool to regulate and observe the function of immune cells that ...

  30. Plasmids 101: Blue-white Screening

    Type
    Blog Post
    ...). A plate with only white colonies is very suspicious! Refrigerate your plates: Placing plates at 4C...α-complementation (cell B). As shown on the representative plate to the right, colonies with an insert-containing...pigment from x-gal (included in the transformation plate medium), turning the bacterial colony blue. We should...backbone plasmid without insert. All colonies on this plate should be blue, indicating that your IPTG and x-... rush the process: It is important to give your plates enough time for any intact β-galactosidase to be... and white colonies. Take care in making your plates: X-gal is light and temperature sensitive and needs...after autoclaving. If spread on top of pre-made plates, make sure it is evenly distributed and allow sufficient...

  31. Viral Vectors 101: Optogenetic Tools

    Type
    Blog Post
    ...that induce conformational changes with light stimulation to either activate or inhibit a specific receptor... receptors  G-protein coupled receptors (GPCR) regulate cellular function through multiple intra-cellular...cryptochrome 2 (CRY2) (Zhang & Cui, 2015). When stimulated with light, the endogenous proteins activate ... a single pathway to activate various protein regulators at different points in the pathway. Photoswitches...with the stimulus. For example, if you want to stimulate at a fast frequency without missing an action ...slower kinetics can be used for lower frequency stimulation. GPCR signaling GPCR signaling is a complicated...considering for your experiment, note that you cannot extrapolate results from one cell type to another, as they...

  32. Technologies Enabled by NanoLuc® Luciferase

    Type
    Blog Post
    ...employing optogenetic regulators to initiate the event being monitored. Optogenetic regulators respond to specific...materials and information. Many of our technology platforms like HaloTag® Fusion Protein, codon-optimized ...infected/expressing cells by flow cytometry. The isolated cells were injected into mice and imaged after.... injection of furimazine or luciferin 24 hours later demonstrated a more than 2-fold brighter signal ...biosensors, the intensity of the FRET signal can be correlated to specific concentrations of biological molecules...initiation wavelengths, excitation of the optogenetic regulator leads to spurious activation of the FRET sensor...biosensors that can even be paired with optogenetic regulators. Many of these tools are available from Addgene...

  33. PiggyBac-ing Through the Genome Editing Field

    Type
    Blog Post
    ...1983, but wasn’t successfully used for genetic manipulation in mammalian cells until 2005 [1]. Like other...stable cell lines is to transfect, select, and isolate clones. This procedure is cumbersome and inefficient... other genetic alterations have occurred) and isolating the specific cells that have the edit of interest... CRISPR/Cas9 to efficiently perform genetic manipulations in animal models or cells lines, including stem...included in a standard homology directed repair template to facilitate the selection of cells containing... technology using a site-specific nuclease to stimulate homologous recombination and Footprint-Free™ removal...re-cutting. Transposase removal of the cassette later restores the target sequence. This allows researchers...

  34. All in a Twist: dsRNA

    Type
    Blog Post
    ...Transcription, translation, and prone to degradation – those are the words that describe RNA! Double ...transcription. In an uninfected cell, dsRNA rarely circulates, but upon viral infection, this nucleic acid ... RNAi is designed against), RISC can repress translation and/or degrade target mRNA. These RNA tools ... almost always epigenetically silenced, but dysregulation of these systems can lead to local dsRNA production...source of dsRNA (that doesn’t involve cellular dysregulation) is the mitochondria. These organelles have ...During transcription, RNA is synthesized from a DNA template, inherently generating RNA:DNA intermediates in...which suppress endogenous dsRNA formation are dysregulated, immune responses can mount in the absence of...

  35. Lighting Up Cell Signaling with Photoswitchable Kinases

    Type
    Blog Post
    ...scientists frequently use growth factors or serum to stimulate a pathway of interest, but there can be a lot ...KTR-mRuby2). The ERK KTR is a target for MEK1 phosphorylation, much like endogenous ERK, and its localization...indicator of MEK1 activity. When MEK1 is active, it phosphorylates the ERK KTR and red fluorescence localizes ...MEK1 and ERK are inactive, the ERK KTR isn’t phosphorylated and red fluorescence can be found in the nucleus...as a stimulus. This system also allows precise modulating of a particular arm of the signaling pathway ...the negative feedback on endogenous MEK1/2 phosphorylation by creating a short pulse of psRaf1 activity...nm light (turn off psRaf1). After this, the phosphorylation status of MEK1/2 was tracked at 5 minute intervals...

  36. A History of Genome Engineering in Popular Culture

    Type
    Blog Post
    ...revolution in genetic engineering techniques is a speculation of yesteryear which has been realized recently...explained how the four letters of the DNA get translated into the twenty amino acids. Once scientists ...DNA encoded all the information necessary for regulating and propagating life, there was no turning back... molecular biologist was to devise methods to manipulate it. The term genetic engineering itself is attributed...what would happen if these corporations would extrapolate genetic engineering to exploit human beings. ...repaired by homologous recombination with a repair template.  Each zinc finger recognizes a nucleotide triplet...advertisement of the technology on various non-scientific platforms, CRISPR has also transcended into the world of...

  37. Common Injection Routes in Mice

    Type
    Blog Post
    ... as “instillation” or “gavage,” as they do not require any punctures. Intratracheal instillation Intratracheal...Skill required: advanced Intranasal instillation Intranasal instillation is very similar to its intratracheal...are used for a broad range of substances, as it circulate what is injected throughout the body. Tail vein...not recommended for tumorigenic compounds, as circulation is slower, and more institutional oversight is...the muscles. The most common injection sites are flat and easily accessible, like on the back. Depending...performed anywhere on the body, somewhere large, flat, and easily accessible (like the back) is preferred...Intratracheal instillation is used to deliver substances directly to the lungs. It is an advanced technique...

  38. Chromoproteins: Colorful Proteins For Molecular Biology Experiments

    Type
    Blog Post
    .... Eight years later, scientists isolated pink, purple, and blue chromoproteins isolated from reef-building...reach zooxanthellae, the coral’s symbiotic dinoflagellates that are photosynthetic (Salih et al., 2000...experiment worked just by looking at the cells on the plate. For example, if your plasmid contains a gene encoding...directly use in bacteria, where most genetic manipulations start. Anthony Forster’s lab used codon optimization... Chromoprotein expression from bacteria on agar plate. Image: Liljeruhm et al., 2018. For each chromoprotein...exchange. These vectors contain a toxin gene (regulated by a rhamnose-inducible promoter) and a chromoprotein...Dove SG, Takabayashi M, Hoegh-Guldberg O (1995) Isolation and Partial Characterization of the Pink and Blue...

  39. Light Sheet Fluorescence Microscopy

    Type
    Blog Post
    ...solution of benzyl alcohol and methyl salicylate which was later modified by others to produce Murray’...major advances in light microscopy, particularly related to the neurosciences.  These developments in microscopy...single gold particles with diameters less than 4nm. Lateral view of a cleared rat pons, medulla and the first...using optical projection tomography (OPT)  (12) and later Hans-Ulrich Dodt’s lab pioneered the use of light...) (6).   The Dodt and Frank Bradke laboratories later developed the 3DISCO (3-dimensional imaging of solvent...tetrahydrofuran-based tissue clearing (13,14). A few years later, Karl Deisseroth’s lab developed CLARITY, which ...relied on the use of transgenic mice in which a subpopulation of neurons are brightly labeled with GFP (e....

  40. Uncovering Ancestral Bias in CRISPR Research

    Type
    Blog Post
    ...tumors, which can lead to new cancer targets and related biomarkers . Data from such screens are often uploaded...Unexpected results Their results showed 96 strongly correlated genetic dependencies that were clearly associated... they looked for a biological explanation, a relationship between the genetic dependencies they identified... ancestries. While there were some exciting relationships identified, 48% were found to be artificial,...bias into their experiments instead of a true correlation.   It turned out there was indeed a bias, and...diversity of genetic variation across the human population. Rather, it primarily reflects European ancestry... to guides in the Broad’s Genomic Perturbation Platform Avana library (used by the Cancer Dependency Map...
Showing: 241 - 280 of 974 results