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We narrowed to 847 results for: PLE

Showing: 281 - 300 of 847 results

  1. Career Coaching for Scientists: Why and Where Do I Find One?

    Type
    Blog Post
    Published
    Jan. 29, 2015, 1:25 p.m.
    ...coaches will have loads of success stories and plenty of people to vouch for them. Trust your gut – only...I have seen, without exception, that this helps people do a better job at reaching their goals.  Check...costs. Can you suggest some coaches? Yes I can. Please note that Addgene does not endorse any of these...Sarah Cardozo Duncan is a proven coach who helps people in science and tech get jobs. She is also a very...? Some coaches focus on graduate students. For example Dora Farkas, founder of Finish Your Thesis (who...based in Spain. The bottom line – there are lots of people out there who want to help. Don’t be afraid to ...

  2. Viral Vectors 101: Calcium Sensors

    Type
    Blog Post
    Published
    Aug. 10, 2023, 1 p.m.
    ...process you would like to monitor? Neurons, for example, have unusually fast calcium dynamics, whereas ... is most sensitive to your expected range. For example, calcium in the ER lumen reaches the sub-millimolar...binding of the biosensor to endogenous calmodulin. Multiplexing Are you combining your calcium reading with ...you need to free up crowded optical space for multiplexing, GECO sensors are available in green, blue, ...Additional red sensors based on mRuby (jRCaMP1) and mApple (jRGECO1a) are also available. Detection methods...single fluorescent protein (FP), requiring only a simple measurement. However, single wavelength measurements...Genetically encoded calcium indicators to probe complex brain circuit dynamics in vivo. Neuroscience Research...

  3. Viral Vectors 101: Chemogenetics

    Type
    Blog Post
    Published
    July 11, 2023, 1:15 p.m.
    ...In order to understand the complex wiring of the brain and the underpinnings of neurological disease,...while resulting in minimal off target effects. For example, if your receptors are expressed in the brain, ...exclusively activated by designer drugs, are G protein-coupled receptors (GPCRs), that have been reprogrammed ...receptor acts through. The DREADD hM3D(Gq), for example, lowers the threshold for neuronal signaling and...themselves are modified ligand binding domains. When coupled to the ion pore domain (IPD) of a channel, the ...expression and signaling of a specifically designed Gi-coupled receptor in transgenic mice. Nature Biotechnology... Multiple Authors...

  4. How to Prepare for an Industry Interview

    Type
    Blog Post
    Published
    July 9, 2020, 1:15 p.m.
    ...task, action, and results. While this method seems simple, I found it hard to remember to describe all four... more than one theme and can be used to answer multiple types of questions. Writing a rough outline of... say You’re going to be interviewed by numerous people during your industry interview. Each interview ...interview. Expect to be tired once the interview is complete Industry interviews are long! Mine lasted 4.5 ...interviewer and then thank them once the interview is complete. Remember, your interviewers took time out of ...remotely Remote interviews are likely new to most people - even hiring managers and your interviewers. You...a question. Maybe you ask for questions after completing each section of your talk. Some video conferencing...

  5. A New Generation of Adenine Base Editors Improves Editing in Primary Human Cells

    Type
    Blog Post
    Published
    May 7, 2020, 1:15 p.m.
    ... or Cas nickase)  a guide RNA that targets the complex of TadA and dCas to the genomic DNA of interest...effect of removing the wild-type TadA from the complex the researchers developed a second set: The ABE8... the team replaced S. pyogenes Cas9 in the ABE complex with PAM-variant Cas9 proteins: the S. pyogenes... ability to use non-standard Cas9 in the ABE8 complexes enables a broader targeting scope for base editing... the corrected cells back to the patients. For example, defects in hemoglobin (𝛽-globin), which cause... is very useful, sometimes, you need to modify multiple genes to generate a cellular phenotype. Again ...that require high DNA editing specificity, for example for potential therapeutic approaches, the authors...

  6. Adenoviral Vector Production and Troubleshooting

    Type
    Blog Post
    Published
    June 14, 2018, 11:44 a.m.
    ...particles = number of particles per mL of sample. The purity of the sample can be estimated by looking at the...negligible toxicity. However, their production is more complex and the presence of helper virus contaminants have...final recombinant adenoviral plasmid construct, please see Addgene’s Adenoviral Guide webpage, and references...which time the cells are scraped, and lysed by multiple freeze-thaw cycles. The resultant lysate is your...amplification.*Pro-Tip*: The cells will become completely confluent and the media will turn yellow. Do ...cytopathic effect.” Amplification can be repeated multiple times (2-4 rounds) at increasing scale over the...

  7. Adapting Toehold Switches to Detect Zika Virus

    Type
    Blog Post
    Published
    Aug. 30, 2016, 2:30 p.m.
    ...high and above what would be expected in patient samples (e.g.: blood, urine). Adapting toehold switches...of the emerging outbreak, as many as 2.2 billion people globally live in areas conducive to the spread ...involved definitely contributed to our ability to complete the project on such a short timescale. Our article...pathogen’s genome as unique barcodes to identify samples containing the virus. Of the 48 sensors, targeting... diagnostic results.  Our approach now detects samples at clinically relevant concentrations (1-3 fM; ... look at viral load in patient blood and urine samples; we showed that we can detect the virus directly...Interview with Previous Collin's Lab Member and Sample 6 Cofounder Michael Koeris Learn How Minigenomes...

  8. Plasmids 101: Terminators and PolyA signals

    Type
    Blog Post
    Published
    March 31, 2016, 2:30 p.m.
    ...structure of the hairpin-DNA complex is thought to destabilize the transcription complex, initiating cleavage ...Polyadenylation is the post-transcriptional additional of multiple adenine (A) nucleotides to the tail of a messenger...polymerase terminates differently. Polymerase III, for example, relies on a specific sequence and RNA secondary...mechanism, whereas Polymerase II termination is more complex and involves two RNA polymerase-associated proteins...polyadenylation enzymes, in a process that seems to couple termination with polyadenlyation. Mammalian expression...PMC104046. Wodrich H, Schambach A, Kräusslich H-G. Multiple copies of the Mason–Pfizer monkey virus constitutive...

  9. Viral Vectors 101: Producing Your rAAV

    Type
    Blog Post
    Published
    July 16, 2024, 1:15 p.m.
    ...single-stranded and self-complementary adeno-associated viral vector genome titers by droplet digital PCR. Human... helper viruses like adenovirus (Ad) or herpes simplex virus to facilitate efficient replication. The ..., and the two helper plasmids, commonly called “triple plasmid transfection” (Figure 1).    Figure 1:...quantitative PCR (qPCR) (Aurnhammer et al., 2012), droplet digital PCR (ddPCR) (Lock et al., 2014), or determination...known titer on every qPCR run. Note that the rAAV sample has to be digested with DNase before undergoing... Multiple Authors...

  10. Pooled Library Amplifications

    Type
    Blog Post
    Published
    May 30, 2024, 1:30 p.m.
    ...library from Addgene, they’ll need to amplify the sample for further downstream applications. As individual...make the library more difficult to work with. For example, a library in a lentiviral backbone might be more... as individual plasmids in your library. As an example, if you want 100x coverage of your library, and...transformation efficiency (Figure 2).    Figure 2: Example of a serial dilution to estimate transformation...libraries that are similar in size and composition as examples. Table 1 provides an overview of pooled library...protocols you can use as a reference. Table 1: Examples of Different Pooled Libraries at Addgene.  ...Libraries (PDF, 401 KB) Screening Zhang Lab - Multiplexed Overexpression of Regulatory Factors (MORF) Library...

  11. Plasmids 101: Degron Tags

    Type
    Blog Post
    Published
    March 14, 2024, 1:15 p.m.
    ...endogenously expressed SCF complex members, an E3 ubiquitin ligase complex. Finally, when auxin is added...fusion protein with the endogenous CRBN E3 ligase complex which will polyubiquitinate the fusion protein....target protein-of-interest and the CRBN E3 ligase complex leading to ubiquitination and proteasomal degradation...acid (IAA), the TIR1 F-box associated E3 ligase complex will ubiquitinate the AID-tagged protein which ... Chung HK, Jacobs CL, Huo Y, Yang J, Krumm SA, Plemper RK, Tsien RY, Lin MZ. Tunable and reversible drug...Inducible, reversible system for the rapid and complete degradation of proteins in mammalian cells. Proc...

  12. Academic vs. Industry Postdocs

    Type
    Blog Post
    Published
    Nov. 13, 2018, 1:22 p.m.
    ...never enjoyed grant-writing. The more I talked to people with industry experience, the more I heard that...decision, the transition felt scary. While I knew two people from graduate school who were at AstraZeneca and...industry for any length of time, you’ll know more people in industry, which means you’re more likely to ...decade of work. The number of steps, decisions and people involved is staggering. The process is complicated...individual could not bring a drug to the market, so people in industry have to work together. Even as a postdoc...postdoc, you’ll have to work with other people in your group, and they are unlikely to hire you if they... academia, promotions in industry are based on people’s ability to manage others and work as a team as...

  13. Addgene's Cancer Collection Pages Connect the Research Community

    Type
    Blog Post
    Published
    Sept. 6, 2016, 2:30 p.m.
    ...millions of new cancer cases arise and millions of people die from cancer-related complications each year...cancer, really? While cancer is extraordinarily complex at the molecular level, at the cellular level there...cancer research has focused on picking apart the complex cellular pathways involved in the development of...pathway. Its major appeal is that it enables the simple construction of vectors to study the function of...transcript most commonly found across all TCGA tumor samples is represented, making the results of any such ...biggest challenges facing researchers today is the complexity of the pathways involved in the development of...continue to to build and develop this new resource! Please leave us your thoughts in the comments section ...

  14. Isolating Genomic Regions of Interest with the CRISPR System

    Type
    Blog Post
    Published
    May 30, 2019, 12:44 p.m.
    ...targeting a specific genomic locus, which can require multiple cloning steps (Fujita et al., 2013). CRISPR-mediated...engineered DNA-binding molecule such as the CRISPR complex is expressed in cells to be analyzed, the Fujii...another "in vitro" form. In this form, the CRISPR complex, consisting of a recombinant dCas9 protein and ...interest with dCas9 and a targeting gRNA. One such example by the Xu lab is CRISPR affinity purification in...purification of a specific genomic region is another example of the many tools based on targeting of a genomic...genomic locus through a specific gRNA-dCas9 complex. When combined with next-generation sequencing, this ...research conducted in the Hodaka Fujii Laboratory, please also see our interview with Dr. Fujii. References...

  15. CRISPR 101: RNA Editing with Cas13

    Type
    Blog Post
    Published
    July 31, 2020, 12:30 p.m.
    ...REPAIR and RESCUE can function in a multiplexed manner to edit multiple RNAs. It’s clear that CRISPR RNA ...research. RNA editing advantages RNA editing has multiple advantages over more traditional DNA editing systems...This change specifies the edit to be made when multiple As are present in the gRNA spacer, as ADAR will...target window, possibly due to longer stretches of duplexed RNA. Using an ADAR2DD catalytic mutant, they showed...By mutating residues in ADAR2 that interact with duplex RNA, they hoped to destabilize ADAR-RNA binding...As discussed above, the ability to edit RNA has multiple advantages, including reversibility and use in...

  16. Capturing a Moment with Marc Zimmer’s “The State of Science”

    Type
    Blog Post
    Published
    July 30, 2020, 1:15 p.m.
    ...years ago. Teaching by example No examination of the state of science would be complete without the inclusion...attempts to answer it in five paragraphs, with examples. If that sounds ambitious, that’s because it is...successful scientists have outgoing personalities to supplement their intellectual prowess and scientific skills...Four, “New Science,” Zimmer tackles more recent examples of scientific advances, and posits the way science... much less familiar with as a biologist--as an example of “big science,” a massive, expensive project ...certainly chapters that could warrant their own books. Multiple books have been written about CRISPR alone, and...

  17. Cpf1: A New Tool for CRISPR Genome Editing

    Type
    Blog Post
    Published
    Oct. 14, 2015, 2:30 p.m.
    ...mediate cleavage rather than the multi-subunit complex employed by class 1 systems. A putative new class...Cpf1 (FnCpf1), they employed an E. coli plasmid depletion assay to discover FnCpf1’s PAM sequence requirements...than following it. The Cpf1 crRNA is also much simpler in structure than Cas9’s; only a short stem-loop...CRISPR systems based on Cas9 were thought to be the simplest CRISPR systems and the easiest to adapt to genome...vectors. In 2017, Zetsche et al. developed a Cpf1 multiplexing approach using a single crRNA array to express...varies widely with gRNA sequence. Both groups used multiple methods to examine Cpf1 off-target editing. First...publication at Addgene. 2. Zetsche, Bernd, et al. "Multiplex Gene Editing by CRISPR–Cpf1 Using a Single crRNA...

  18. Plasmids 101: How to Name Your Plasmid in 3 Easy Steps

    Type
    Blog Post
    Published
    Feb. 13, 2014, 1 p.m.
    ...good guidelines to follow in order to ensure that people can quickly and easily identify what your plasmid... empty backbone name in your plasmid name. This simple piece of information can often convey many important...insert abbreviation to specify what species it is. Example: ‘h’ is for Human (homo sapiens), ‘m’ is for mouse...plasmid and their relative position to the insert. Example, if you have a Flag tag on the N-terminal of your...directly before or directly after the insert name.   Example: If the Glutamine at position 295 was mutated to...pBACKBONE-Flag-hGene-EGFP or pBACKBONE-Flag-hGene(wt)-EGFP These simple rules allow any scientist to know what is in a ...

  19. Hot Plasmids: Fall 2024

    Type
    Blog Post
    Published
    Nov. 21, 2024, 1:30 p.m.
    ...of CRBNmidi alone (A) and in a ternary complex with the multiple myeloma drug mezigdomide and ubiquitination...Construction of solid matrix-antibody-antigen complexes containing simian immunodeficiency virus p27 using...on its own, with drug ligands, and in ternary complexes with other proteins, establishing CRBNmidi as ...mutations.  Here, the researchers were able to target multiple genes in M. abscessus either simultaneously or... (also cyan) embedded in the Titin I10 protein (purple) only when Titin I10 unfolds under tension. Upon... Multiple Authors...

  20. Plasmids 101: Broad Host Range Plasmids

    Type
    Blog Post
    Published
    April 11, 2024, 1:15 p.m.
    ...maintain itself within host cells.  The plasmid has multiple ori that are functionally differentiated from ...proteins from bacterial hosts. In a plasmid with multiple oris, one may function in one type of host and... the others are functional in other hosts. For example, the pJD4 plasmid harbors distinct origins (ori1... load (Meyer 2009). Figure 1: Idealized examples of a narrow host range plasmid with only one ori... ori and a broad host range plasmid with multiple oris that vary in DnaA-box and iteron structure, host-independent...Acinetobacter calcoaceticus, Actinobacillus pleuropneumoniae, Actinomyces naeslundii, A. viscosus, Aerobacter... hosts in which proteins can be expressed. For example, scientists can incorporate interchangeable antibiotic...
Showing: 281 - 300 of 847 results