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We narrowed to 1,000 results for: Eras

Showing: 261 - 280 of 1000 results

  1. Five Popular Model Organisms

    Type
    Blog Post
    ...so scientists can simultaneously observe several generations of mice at a time. Because mice are more ...better recapitulate the complex interactions between cancer cells, therapeutic drugs, and the rest of the ... easily maintained in the lab, and have short generation times that make it easy to study the effects ...for scientists as they have a relatively short generation time for mammals - the time between being born...successfully helped researchers develop a variety of therapies. For example, many scientists use mice to study...mutations that made cancer cells more resistant to chemotherapy. You can find these plasmids at Addgene. Another...important human disease model with potential for therapeutic drug discovery as it is estimated that 75% of...

  2. Technical Design of a Western Blot

    Type
    Blog Post
    ...your target! Incubation temperature and time Incubating at a lower temperature reduces background noise...options are 4 °C overnight or 1–2 hours at room temperature (RT). Most people choose to incubate their primary...their secondary antibody for 1–2 hrs at room temperature, which allows them to reduce background while...for any available data on the best incubation temperature for your specific protein(s) of interest.  Determining...shelf life; runs at neutral pH (reduced protein alteration) Expensive buffers Bis-tris 6–400 kDa...shelf life; runs at neutral pH (reduced protein alteration) Expensive buffers Tris-tricine 2.5–40...gel. Handcast gels are significantly cheaper and generate less waste. However, they also add time to an ...

  3. Plasmids 101: Gateway Cloning

    Type
    Blog Post
    ...mammalian cells. STEP 1: Generate an Entry Clone There are a few different ways to generate our desired entry...interest to generate entry clones Entry vector attL sites for recombination Used to generate entry clones... phage from the bacterial chromosome and the regeneration of attP and attB sites.   Figure 1: ...is catalyzed by the BP Clonase enzyme mix and generates the entry clone containing the DNA of interest...Reaction takes place between the attL sites of the generated entry clone and the attR sites of the destination... the DNA of interest flanked by attB sites is generated. As in the BP reaction, a DNA fragment containing...experiment where we might use Gateway cloning to generate our desired constructs: lentiviral expression ...

  4. Tips for Using FRET in Your Experiments

    Type
    Blog Post
    ...molecule interactions in living cells using a microscope. Historically, molecular interactions were detected...first to be used to study protein-protein interactions and several other pairs have been used since - including...technique which allows the detection of molecules' interactions, modifications or dissociations in situ. Used...often using probes with the potential to target several molecules. By analogy, it was like pointing out...hall but not knowing if these students know or interact with each other. FRET reduced the scale of our...our perception about molecular interactions. What is FRET? In their JCB 2003 paper, Sekar and Periasamy...measurement sensitivity makes it suitable for studying interactions within living cells. By coupling fluorophores...

  5. Stem Cell Models for Disease & Open Science: Interview with Darrell Kotton

    Type
    Blog Post
    ...multidisciplinary nature of the Center for Regenerative Medicine accelerates research. Chapter 5: Research currently... in therapy. The lungs are difficult, but other tissues like the retina have cell-based therapies underway...Darrell Kotton is the director of the Center for Regenerative Medicine at Boston University Medical School...source biology in his own lab, at the Center for Regenerative Medicine, and within the biomedical research... Kotton and the Boston University Center for Regenerative Medicine. Darrell describes his career goals...researcher. Darrell's ultimate goal - to develop therapies to treat disease in the lung. Chapter 2: Balancing...discovery of GFP. Chapter 4: Goals of the Center for Regenerative Meidince - Understanding and developing means...

  6. Supporting Rare Disease Research with the 2020 BeHEARD Awards

    Type
    Blog Post
    ...Mark Mellett: IL-36 biology in generalized pustular psoriasis Generalized pustular psoriasis is a chronic...autoinflammatory loop: keratinocytes both respond to and release IL-36 cytokines. Keratinocytes are the main source...36 release in keratinocytes,” says Mellett. This would help the lab identify therapeutic targets in the...Genomics Institute BeHEARD (Helping Empower and Accelerate Research Discoveries) Challenge provides grants...is powered by ATP. A molecule central to the regeneration of ATP from ADP is creatine. In order to reach...could greatly enhance our ability towards CTD therapeutic development,” says Iyer. Since the RESOLUTE plasmid...cytokine family resulting in the activation of keratinocytes, which release inflammatory mediators that trigger...

  7. Hot Biosensors 2022: Year-End Roundup

    Type
    Blog Post
    ...capable of local temperature sensing with sub-millisecond resolution and sub-degree temperature differences...Figure 7: B-gTEMP temperature-sensitive probe enables fast measurement of temperature and local heat transfer...other targets. Figure 1: MTRIA biosensors are generated by inserting a circularly-permuted GFP into an...localized for synaptic glutamate response. They generated variants optimized for different applications ...phosphorylated. To demonstrate that pYtags can be generalized to other RTKs of interest, they create sensors...deep-tissue imaging. Vladislav Verkhusha’s lab has generated a new genetically encoded calcium indicator using...affinity isn’t always better, especially if it means operating at the outer edge of the sensor’s dynamic range...

  8. Five Popular Model Organisms, Part 2

    Type
    Blog Post
    ...for its incredible regeneration abilities. The axolotl has the ability to regenerate complex structures..., 2019) Besides their ability to regenerate, the axolotl has several attractive characteristics that make...genes related to limb regeneration (Vieira et al., 2019). To study regeneration scientists have created...Because E. coli are easily propagated and have a generation time of around 20 minutes, they are perfect for...been used to identify and study protein-protein interactions in proximity labeling techniques such as BioID...processes and signaling pathways that regulate regeneration and wound healing are conserved across species...potential humans have for a similar degree of regeneration.  In addition, axolotls have many age-related...

  9. Getting Started with Behavioral Neuroscience: Standardizing Experiments and Using Viral Vectors

    Type
    Blog Post
    ...experimental design considerations used in all behavioral studies. General considerations for behavioral ...behavioral testing As a general consideration for behavioral testing in a laboratory, a high level of experimental...It is a very old branch of science, dating back several hundred years, with historical ties to psychology...given some time to adapt to a testing apparatus. General stress levels of the test animal may also be reduced...handling for a few minutes by the experimenter several days preceding the experiment. Handling can however...mouse breeding colony) Environment Olfaction, temperature, humidity, noise level, circadian effects (inactive...inactive versus active phase) Experimenter Interaction should be minimal and strictly controlled; optional...

  10. CRISPR 101: Drug discovery and CRISPR

    Type
    Blog Post
    ...candidate genes! Disease model generation CRISPR has made the generation of genome edits accessible, fast...select and generate physiologically relevant models. In complex diseases, it can aid in generation of multiple...respond to an existing therapy. In theory, CRISPR could even be used to generate patient mutations and ...cancer and identified several candidate inhibitors of gene X in a screen. You generate a gene X knockout ...design. Alternatively, a disease model can be generated, and drugs screened for a change in pathology....identification of potential targets by CRISPR can accelerate the drug pipeline. High-throughput CRISPR screens..., gene knock out, etc. – CRISPR can be used to generate cell line or animal models which have the precise...

  11. Meet the New Addgene Board Members!

    Type
    Blog Post
    ...employees at Dimension Therapeutics, she helped build and grow the gene therapy division of the company...journal Molecular Therapy. Vandenberge is also the Grousbeck Family Chair in Gene Therapy, Director of the...Khandan Baradaran’s expertise in viral safety, gene therapy vector development, vaccine development, and assay...Addgene. She was Head of Quality at Dimension Therapeutics, which was acquired by Ultragenyx Pharmaceutical...Baradaran met Addgene Executive Director Joanne Kamens several years ago through the Healthcare Businesswomen’...Stegmeier is the Chief Scientific Officer at KSQ Therapeutics and is an Addgene depositor. KSQ uses a CRISPR-Cas9...the first lentiviral microRNA-based system for polymerase II-regulated RNA interference. In the first two...

  12. Using AAV for Neuronal Tracing

    Type
    Blog Post
    ...and anterograde transport suggest that there are several parallel mechanisms in place (for review see Maday...retrograde tracing, albumin protein labelled with horseradish peroxidase (HRP; Kristensson and Olsson, 1971... Virus (RABV) which moves retrogradely from a peripheral site of infection to the central nervous system...it can replicate and by doing so spread across several synaptic connections (Ugolini et al., 1989). When...Lo & Anderson, 2011). In the past two decades, several virus species have been modified and used as tools... the viral proteins VP1, VP2, and VP3. To date several hundred variants, or serotypes, have been found...et al., 2008; Watakabe et al., 2015). There are several reports of both retrograde and anterograde intracellular...

  13. Predicting Adverse Reactions to Monoclonal Antibody Drugs

    Type
    Blog Post
    ...Monoclonal antibody drugs are popular therapeutics for a plethora of disease conditions, from cancer ... they elicit an immune response from the body. Several factors contribute to the immunogenicity of a drug...response. T cell epitopes are found in either the therapeutic antibody itself or as impurities in the formulation... algorithm to evaluate the immunogenicity of therapeutics in silico. These algorithms are trained on vast...score (Mattei et.al., 2022).    Figure 1: Generation of immunogenicity scores by predictive algorithms...algorithms fail to capture. In vitro evaluation of ‌therapeutic immunogenicity is typically performed using traditional...traditional 2D assays with immune cell lines or peripheral blood mononuclear cells (PBMC) in 3D models of...

  14. Using CRISPR/Cas9 to Edit Disease Out of the Genome

    Type
    Blog Post
    ...Advances in CRISPR/Cas-based Gene Therapy in Human Genetic Diseases.” Theranostics. https://doi.org/10.7150/thno...possibility to treat human genetic diseases, and several treatments are currently being tested in clinical...discovery/development of genetic targets and RNA therapies. [6]. In 2014, we wrote a blog post on CRISPR’...researchers are now using it to investigate potential therapeutics for complex genetic diseases. So we’re revisiting... showed that CRISPR/Cas9 could be used to edit several autosomal-dominant mutations that have been connected...s team noted that CRISPR/Cas9 has been used to generate mutations in a wide range of organisms, but its...stem cells in organoid cultures are stable,” said Gerald Schwank, first author of the cystic fibrosis report...

  15. Plasmids 101: Sequence and Ligation Independent Cloning (SLIC)

    Type
    Blog Post
    ...The key is T4 DNA polymerase, which has 3′→5′ exonuclease activity. T4’s polymerase activity can be thwarted...For one thing, those 5′ overhangs can be generated in several ways. One option, imprecise T4 digestion...the target vector. 5′ overhangs can be generated by T4 polymerase (left branch); by annealing two separate...work directly on PCR products, with 5′ overhangs generated either by annealing a mixture of products or from...concentrations of insert to succeed. 5′ overhangs are generated in the linearized vector by T4 digestion; then...adjacent fragment or vector. 5′ overhangs are generated on all the fragments and the vector, specifying... this limitation, one option is to perform a hierarchical assembly, assembling fragments in multiple steps...

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

    Type
    Blog Post
    ...basic immunology, cancer-immune cell interactions, cancer immunotherapy, as well as new approaches to reprogramming...tumor-associated inflammation for therapeutic benefit ”as stated by the organizers. My overall impression from this... discussed the effects of starvation on cancer therapy. Findings in previous animal studies suggested ...immunosuppressive reaction in order to improve therapeutic outcomes.  If you are studying inflammation and...visualization techniques it is possible to follow the interactions between tumor and immune cells in real time.... UCSF showed some impressive videos of these interactions (Ref 3). You can find useful tips on in vivo...to help rid us of cancer in combination with chemotherapy. To dive deeper into this field read our recent...

  17. All in a Twist: dsRNA

    Type
    Blog Post
    ..., such as chemotherapy treatment or ionizing radiation, can also result in dsRNA generation. While these... Here, we will cover how these structures are generated, why it’s important for cells to sense their formation...these viruses have one thing in common – they can generate dsRNA as part of their infection and/or replication...inherently double stranded, while ssRNA viruses can generate dsRNA intermediates as part of the viral replication...mRNA. These RNA tools have been used in clinical therapy to target pathogenic proteins, as well as in the...as long/short interspersed nuclear elements can generate dsRNA by bidirectional transcription or as inverted...is synthesized from a DNA template, inherently generating RNA:DNA intermediates in the process. These transcription-coupled...

  18. CRISPR 101: Cas9 vs. The Other Cas(s)

    Type
    Blog Post
    ...complementary site in the genome where it will proceed to generate a blunt DNA double strand break. Cas9 has been...Cas12’s primary difference from Cas9 is that it generates staggered breaks, also known as sticky ends. Cas12a...targeting experiments requiring single strand DNA generation, such as homology directed-repair. Cas12a is ...outlined below as well. Cas12a also has significant therapeutic promise due to the fact that it is smaller than...simplicity of Cas12a makes it easier to package for therapeutic delivery by AAV, which has strict capacity limitations...sequences of Cas12a to further optimize it for therapeutic and laboratory usage. Fast facts – PAM requirement...TTTN. Best for: homology-directed repair or therapeutic delivery.   Cas14 – Mini, but single strand ...

  19. Working with Nuclear Receptors

    Type
    Blog Post
    ...such as p300 and CBP histone acetyltransferases or CARM1 methyltransferase. This attendant signaling feature...years evolved from chloramphenicol acetyltransferase to luciferase, the basic assay remains widely used...Both physiological processes are controlled by deliberate and compulsory actions instigated by the central...central nervous system signaling to peripheral effector organs, which then return information such as nutritional...communication of such signals between central and peripheral organs because of their established roles as ...available or being developed to treat a multitude of therapeutic indices, including hypertension, cancer, diabetes...action). Primary or core coregulators (those that interact directly with NRs) exist in steady-state complexes...

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

    Type
    Blog Post
    ...challenge for CRISPR HIV-1 therapies is sequence specificity. Whereas antiviral therapies target HIV-1 at the...Today, HIV-1 can be controlled with antiviral therapies such that the virus is undetectable in the blood...demonstrated two potential strategies for CRISPR-HIV therapeutics - one using dCas9-SAM to activate HIV-1 transcription...usually seen in healthy individuals. Antiretroviral therapy (ART) can virtually eliminate plasma HIV-1, improving...binding sites increased activation of HIV promoter-luciferase constructs. In multiple HIV-1 latent T cell lines...the probability of off-target effects. In such a therapy, CRISPR/Cas9 would need to 1. excise the HIV-1 ...most HIV-1 patients. Obstacles for CRISPR HIV-1 therapy Both of these methods represent exciting advances...
Showing: 261 - 280 of 1000 results