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We narrowed to 922 results for: Por

Showing: 391 - 420 of 922 results

  1. Plasmids 101: Inducible Promoters

    Type
    Blog Post
    ... phage promoter pR, preventing expression of a reporter gene. When light is present, YFI is inactive, ...preventing repressor cI synthesis and allowing reporter gene transcription to take place. Addgene also...promoter is right for me? Here’s a list of some important variables to consider when choosing an inducible...’re expressing a gene that may be toxic, it’s important that your inducible promoter not be too leaky....

  3. Building and Validating Recombinant Antibodies: A Case Study

    Type
    Blog Post
    ...about the process that was used to create this important collection.  Creating the Plasmids To begin constructing...antibody (R-mAb) expression plasmids, allowing for important reagents that could have been lost to continue...sequences available from each plasmid webpage. Importantly, the methods developed by the Trimmer Lab for...why choosing the right secondary is of primary importance. PLoS One. 2012;7(6):e38313. doi: 10.1371/journal.pone...

  4. Viral Vectors 101: Viruses as Biological Tools

    Type
    Blog Post
    ...permanently integrate into the host genome or temporarily express whatever it encodes. This ability to ...straightforward across applications, while electroporation of a human, for example, isn’t a reasonable...allows you to infect cells and deliver cargo, it’s important to understand their differences to choose the ...PMC413431. 2. Bukrinsky MI, et al. “Active nuclear import of human immunodeficiency virus type 1 preintegration...

  5. Developing a cold tolerant E. coli using specialized chaperones

    Type
    Blog Post
    ...severely cold conditions. To understand their unique importance, let’s begin with a few basics.  Mesophiles vs...understanding and appreciating their biology. The importance of psychrophilic enzymes From an industrial perspective...decided to create a hybrid system where we could incorporate cold-tolerance inducing genes within a regular...scientist from Addgene. The talk highlighted the importance of reagent repositories and the myriad facilities...

  6. Opto-Nanobodies: Using Light to Manipulate Cell Signaling and Protein Purification

    Type
    Blog Post
    ...binding ability of its nanobody partner, so it’s important to insert AsLOV2 at a spot in the nanobody that...light to control signaling provides greater spatiotemporal resolution than pharmacological or genetic ...activated Ras/Erk signaling by using a fluorescent reporter for Erk kinase translocation. These results demonstrate...control protein activity Read about RANbodies, reporter nanobody fusions Learn more about using the secondary...

  7. Choosing a CRISPR Nuclease: Site Accessibility, Specificity, and Sensitivity

    Type
    Blog Post
    ...nuclease for your experiments, there are three important factors to consider: site accessibility, specificity...potential for off-target cleavage should be an important consideration during any experiment. Sensitivity...stringent PAM specificity. This can be especially important for base editing techniques, where Cas9 must be...many options! Comic by Maya Kostman. Another important consideration when choosing a Cas9 is balancing...

  8. How to Make Friends and Meet People at a Scientific Conference

    Type
    Blog Post
    ...scientific conferences. Conferences provide the opportunity to meet people who are interested in the same...the conference. Large conferences have many opportunities but can be overwhelming as far as meeting new...the conference The conference may start in the airport. If you see someone traveling with a poster roll...friendly so stick with those. Don’t miss social opportunities, but don’t overdo it (some of my best studies...

  9. 3 Challenges in Plant Synthetic Biology

    Type
    Blog Post
    ...plants are the foundation of life on earth, and opportunities abound to build better fuels, feeds, foods, ...areas in plant biotechnology that are ripe with opportunity. We’ve decided to jump into one of those unexplored...out their mission of creating new forums and opportunities for open innovation in plant synthetic biology...build the plant synthetic biology community and to support more efficient engineering of a better tomorrow...

  10. Form Your Own Peer Mentoring Group: A How-To Guide for Scientists

    Type
    Blog Post
    .... But how do you find these elusive teachers, supporters and advisors? I tried to start a more formal ...for their development plans. They all found the support of the group to be intrinsic to their success. ... continue to meet regularly, providing ongoing support, advice and accountability for development goals...mentoring relationship.  Celebrate your successes and support each other as you transition to the next stage ...

  11. Seven Tips for Using LinkedIn as a Scientist

    Type
    Blog Post
    ...people can’t find you on LinkedIn, you will lose opportunities and hiring managers will think it is odd. For... you don’t add a personal note, you waste an opportunity to start things out right. Also, many people ...requests without a personal message and they may report you as spam or “I Don’t Know This Person.” If that...had a chance to proof read. This might not be important for a good friend, but if you are making a professional...

  12. Plasmid Tools for Microbiome Studies

    Type
    Blog Post
    ...microbiome by using engineered genetic tools and reporter microbes. Let’s take a look at a few of these ... because in vitro conditions did not optimally support growth of diverse species. These results further...is a ribosome binding site (RBS) followed by a reporter gene. The hairpin blocks translation by sequestering... reveals the RBS and allows translation of the reporter gene resulting in GFP signal. With the addition...

  13. Hot Plasmids - March 2019 - Anti-CRISPR, 2in1 Cloning, Fluorescent Voltage Indicators, and Photoswitchable Proteins

    Type
    Blog Post
    ...2017). However, Acr inhibition is spatially and temporally ubiquitous, making them a rather blunt instrument...versatility opens up the potential for spatial and temporal control in many genetic targeting and editing ...rapidly screen thousands of proteins for one that can report electrical activity via imaging. They took the ...response to light, offers a way to spatially and temporally control cellular processes in vivo. Many photoactive...

  14. Human Germline Editing Using CRISPR

    Type
    Blog Post
    ...containing Cas9 and two different gRNAs, and electroporated them with an ssODN repair template in induced...in cases of disease prevention. However, it is important to consider that already-tested CRISPR alternatives...clinical applications could be considered. It is important to consider the question of broader applicability... large step forward in the CRISPR field, it’s important to remember that the study hasn’t achieved any...

  15. Rosella: A Fluorescent pH-Biosensor for Studying Autophagy

    Type
    Blog Post
    ...spectra. DsRed is the permanent fluorescent tag portion of Rosella: it will emit red fluorescence regardless...mechanisms behind autophagy by tracking what’s being transported to the yeast vacuole (i.e. cytosol, mitochondria...2008). Rosella: A fluorescent pH-biosensor for reporting vacuolar turnover of cytosol and organelles in...using a single fluorescent biosensor. Scientific Reports,5(1). doi:10.1038/srep12397. PubMed PMID: 26215030...

  16. Giving gRNAs a Facelift - Synthetic and Beyond

    Type
    Blog Post
    ...Nature has evolved ways to prevent degradation of important cellular RNA molecules with post translational...your gRNA. Pro tip: modifications are typically incorporated at the termini of the crRNA molecule (usually...Frontiers in Genome Editing). This is particularly important in experiments in which viability is being compromised...Ramel, MC., et al. Photoswitchable gRNAs for Spatiotemporally Controlled CRSIPR-Cas-based Genomic Regulation...

  17. Overcoming the Challenges of Lentiviral Production

    Type
    Blog Post
    ...viral Rev gene from the packaging plasmid and incorporating it into a fourth plasmid. In addition, the promoter...third generation packaging systems, this system incorporates the Tet-off transcriptional activation system...number of harvests will all impact the titer. Importantly, optimized conditions for one transfer plasmid...producing lentivirus, it is time to titer. One important consideration when titering a lentivirus is the...

  18. Open Resources and Plasmid Tools For Studying C. elegans

    Type
    Blog Post
    ...publication.   3’UTRome The 3’UTRs of mRNA are important for post-transcriptional regulation in C. elegans...combination of these methods provides unique opportunities to develop computational methods to better understand...system allows scientists to both spatially and temporally knock-down proteins of interest.  Plasmids for...intersectional strategy using a split intein for refined spatiotemporal transgene control in Caenorhabditis elegans...

  19. Design Tips for Prime Editing

    Type
    Blog Post
    ...that meant rereading many of the original papers reporting various prime editing tools. These papers are ...editing application. And optimizing its sequence is important, as prime editing efficiency is affected by the...nucleotides. For longer templates, it is even more important to test different lengths and sequences, as unintended... cellular surveillance. To limit unintended incorporation of the pegRNA scaffold sequence into the genome...

  20. Antibodies 101: Stripping and Reprobing Western Blots

    Type
    Blog Post
    ...directly to buffer without a re-wetting step. Important: do not dry your membrane with antibodies on it...the ones shared by Bio-Techne, ThermoFisher, or Millipore Sigma. We’ll share the basic composition of these...Alternatively, companies including ThermoFisher, Millipore Sigma, and Azure Biosystems offer pre-made stripping...your western blot several times — some researchers report up to ten cycles! Just remember that a bit of sample...

  21. Hot Plasmids February 2024

    Type
    Blog Post
    ...difficult to genetically edit due to toxicity from electroporation or poor cellular entry with other transfection...PAGE editing with viral sgRNA targeting mCherry reporter (mChe), with co-incubation with assist peptide.... CAP-Mac also robustly expressed fluorescent reporters in green monkeys and cultured human neurons, with...slightly reduced efficacy compared to AA9-X1.1. Importantly, AAV1-X1 is not neutralized by pre-existing antibodies...

  22. Hot Plasmids and Viral Preps - March 2021

    Type
    Blog Post
    ...length can and should be varied as it can have important implications for the fusion protein effectivity...upon these original tools that allows for both spatiotemporal protein degradation and quantification of this...et al., 2021.   The AID system relies on a transport inhibitor response 1 (TIR1) expressed under a ...a desired promoter with a specific spatiotemporal expression pattern and an auxin inducible degron (AID...

  23. Hot Plasmids and Viral Preps - January 2021

    Type
    Blog Post
    ...in a protein.    Cells expressing an RFP reporter plasmid for recombinase activity were exposed ...interest. However, FACS is unable to spatially and temporally target single cells. Now, Francois St-Pierre’...way to isolate individual cells with unique spatiotemporal phenotypes from large heterogeneous cultures...introduction & resolution of two DSBs in a fluorescent reporter. Find the plasmids here Yongsub Kim’s lab engineered...

  24. Fluorescent Proteins 101: Fluorescent Protein Timers

    Type
    Blog Post
    ...have been around since 2000 when Terskikh et al. reported the production of dsRed E5. This timer predictably... your experimental setup less cumbersome. 3. Portability across multiple systems Terskikh et al. showed...fluorescent timers that change color from blue to red report on cellular trafficking." Nature chemical biology...Chronology of islet differentiation revealed by temporal cell labeling." Diabetes 58.8 (2009): 1863-1868...

  25. Degrading DNA with Cascade-Cas3

    Type
    Blog Post
    ....com.   The Cascade complex Cascade is an important component of the CRISPR Cas3 system. Most Cascade... appears to be conserved. This DNA degradation portion can be bypassed by leaving out Cas3 — a tactic ...is pulled through the Cas3. This mechanism is important for Cas3’s biggest trick. Cas3 likes to bluff,... — Cas3 will definitively degrade the targeted portion of DNA using only a single gRNA. Cas3 appears to...

  26. Viral Vectors 101: Voltage Indicators

    Type
    Blog Post
    ...sensors are better classified by their fluorescent reporter and the mechanism of their voltage sensing component...rhodopsin proteins were initially evolved for ion transport, not fluorescence, they are not nearly as bright...for penetrating deeper tissues depths (1 mm), important for certain applications such as in vivo tissue...rhodopsin Ace2N and mScarlet. Ace2N quenches a proportion of mScarlet's emission via FRET.  Image from ...

  27. PhD Applications After COVID

    Type
    Blog Post
    ...in-person portion got canceled, taking advantage of virtual meetings can still be a great opportunity to introduce...course of the last few years: recognizing the importance and benefit of remote and video communication...staff have become a new way to allow you to get important information about the campus even if you can’t...

  28. 3D Printing Meets CRISPR Cas9

    Type
    Blog Post
    ...this rarely happens. But if you ever have an opportunity to read one of these structure papers – while...function in the human body?” … and probably most importantly… "how do we know this?” Over the past fifteen... nuclease active site wasn’t even close to its reported cleavage site on the complementary DNA strand....model also displays the several Lys/Ala mutations reported by Ian Slaymaker and colleagues to reduce off-...

  29. Truncated gRNAs for Regulating Gene Expression

    Type
    Blog Post
    ...suffers from several limitations. Perhaps most importantly, most Cas9 orthologs are less well characterized...(1, 3). The gRNA can be further modified by incorporation of an RNA hairpin, such as the MS2 hairpin, ...with nuclease-competent Cas9. An RNA hairpin incorporated into the truncated gRNA recruits a transcriptional...target site. Benefits of truncated gRNAs Importantly, the use of truncated guides has largely been...

  30. SciComm: Taking Science to Elementary and Middle Schools

    Type
    Blog Post
    ...has made strong efforts in the last 5 years to incorporate more science and technology into the school curriculum...presentation to their students. I didn’t think of this opportunity as coming from networking at the time, but that...presentation because, in my opinion, it was more important to communicate the applications of CRISPR over...use it? This last section was arguably the most important, and it was a little worrisome for me since bioethics...
Showing: 391 - 420 of 922 results