We narrowed to 846 results for: PLE
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TypeBlog PostPublishedFeb. 9, 2023, 2:15 p.m....When Karl Barry Sharpless was awarded this year’s Nobel Prize in Chemistry, he joined an elite club, ...his program. Initially, Provost made his labs completely open-ended, allowing every student to work on... the culmination of many such experiences. For example, students in Provost’s course recently used a combination...Kowalski et al., 2016; Scott, 2021). In fact, completion of Provost’s CURE increased students’ confidence...Community (MCC) that seeks to reduce barriers to implementation. Funded by the NSF (with Co-PIs Jessica Bell...for your classroom, I highly recommend you try implementing one. It's likely only a matter of time before...Poirier, V., Pellettieri, J. 2020. The exon junction complex is required for stem and progenitor cell maintenance...
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Better Dyeing Through Chemistry & Small Molecule Fluorophores
TypeBlog PostPublishedSept. 8, 2016, 2:30 p.m....removing excess dye from the sample, which is particularly important for samples where washing is difficult...reason is something called “the photon budget”: each sample has a limited number of fluorophores and each fluorophore...of information one can extract from a biological sample is wholly dependent on the photon budget and pushing...fluorescence microscopy often requires more photons. For example, moving from transient overexpression of protein... desperate to extract a few more photons from a sample. Chemical fluorophores can be substantially brighter...labeling chemistry easier and more functional in complex biological environments such as live cells and ...brightness of the resulting conjugate, and the complexity of the system. A particularly desirable property... -
Interview: Hodaka Fujii on enChIP, New CRISPR Tools, and More
TypeBlog PostPublishedDec. 2, 2014, 7:23 p.m.... for example, helping to improve the papers submitted to journals. In addition, it is my pleasure to interact...enChIP system enables us to express the CRISPR complex much easier in various cells, including primary...technologies consisting of iChIP and enChIP are recent examples. We also developed the inducible translocation...and PLoS One. Additionally, you are involved in multiple professional scientific societies. Why is being...system of journals and grant applications, for example. However, at the same time, it's fun to be involved...give to post-docs and graduate students as they complete their appointments in your lab? Fujii: These ...develop new technologies, hopefully they'll be completely different from what we are now doing in my lab... -
Starter Guide to induced Pluripotent Stem Cells (iPSCs) Part 2: Reprogramming and Transdifferentiation
TypeBlog PostPublishedNov. 6, 2018, 1:12 p.m....reprogramming factors can be delivered and expressed in multiple somatic cells via various methods. Some of the...directed differentiation.” Specific cell media, supplements, bioactive small molecules, and growth factors...natural transdifferentiation is rare in mammals, an example is observed in the pancreas when excess β-cell ...1987, Davis et al. reported one of the earliest examples of transdifferentiation in vitro where treatment... of interest for in vivo transdifferentiation. Multiple cell types such as fibroblasts, hepatocytes, and...vitro Both in vitro and in vivo Epigenetic marks Complete removal to convert to iPSCs and re-establishment...major processes by which cells are converted to replenish and repair cells that are lost or damaged. Both... -
Neuronal labeling with Spaghetti Monster
TypeBlog PostPublishedAug. 14, 2018, 12:42 p.m.... The central nervous system (CNS) orchestrates complex processes enabling organisms to control their movements...the visible spectrum, researchers can also use multiple FPs in single experiments and visualize numerous.... smFPs are literally composed of FPs fused to multiple epitope tags. Indeed, to create the smFPs Looger...labeling of these fine structures through their multiple, high-affinity binding sites for primary antibodies...expressed and/or similar proteins Through their multiple, highly specific antibody binding sites and primary...used to distinguish highly similar proteins. For example, N-cadherin (cadherin-2) is a postsynaptic cell...super-resolution STORM imaging, and electron microscopy. Sample preparation for these techniques sometimes makes... -
CRISPR 101: Cytosine Transversion Editors
TypeBlog PostPublishedMarch 25, 2025, 1:15 p.m....and replaced it with a double-bonded oxygen. This simple enzymatic reaction enabled conversion between the...and one-ring pyrimidine base. This is a far more complex chemical task, not a job for a single enzyme. ...bind DNA, Cas9 requires (1) a guide RNA, (2) a complementary DNA sequence, and (3) a protospacer-adjacent...that site is filled depends on cellular factors. Multiple labs tried to identify beneficial factors to include...separate and preventing repair based on the complementary strand. This could explain why the edited U ...original G is not primarily converted back to a complementary C. Optimized base editing with eOPTI-CGBE and...so the Yidi Sun and Erwei Zuo labs tested and implemented several of these advances for CGBEs (Yuan et ... -
Tips for Writing a Good Cover Letter
TypeBlog PostPublishedMay 21, 2024, 1:30 p.m....specific examples. Instead of saying “I have excellent communication skills” try giving an example, like ...application process, it's a good idea to talk to people in the field to understand what the norms are. ...match the job requirements, by giving specific examples of your work and explaining how your experience...pointing to your transferable skills. In the above example, if you don’t have experience with ddPCR, but do...beginning is often the hardest part, here are some examples to get you started! Dear Dr./Professor [last ...strengthened an application. Even so, there were examples of great hires who hadn’t submitted a cover letter... Multiple Authors... -
CRISPR Activation: A Practical Guide
TypeBlog PostPublishedAug. 18, 2016, 2:30 p.m....certain loci to create changes in phenotypes. For example, CRISPR activation has been used to differentiate...gRNAs targeting every gene in the human genome for example, can be made easily and cheaply using oligo library...allows for exploratory-based research as well. For example, CRISPRa can be used to find potential cancer drug...et al., 2019). Because of its ability to target multiple gene loci simultaneously, CRISPRa can also be ...(Chavez et al., 2015)) are good choices across multiple cell lines (HEK293T, MCF7, U2-OS, Hela, N2A, 3T3...Church G (2016) Comparison of Cas9 activators in multiple species. Nat Methods 13:563–567 . https://doi....transcriptional activation by an engineered CRISPR-Cas9 complex. Nature 517:583–588 . https://doi.org/10.1038/nature14136... -
Data Freedom: The Expansion of Data Sharing in Research Publications
TypeBlog PostPublishedAug. 5, 2014, 6:51 p.m....of storage. For example, mouse behavioral geneticists may track the motion of multiple animals on different...limiting numbers of figures such that many have multiple, complex panels. PLOS, itself, has not finalized its...Sharing Policies, etc.) and is founded on the principle that scientific publications must contain enough...some data, etc. The firestorm that followed implementation of the policy comprised of numerous objections... paper are in scope. For images, the number of examples supporting the published material is a factor ...collections and interpretation of data, publishing complete data, rather than just a single projection of ... -
Scientist Networking: What is an Informational Interview?
TypeBlog PostPublishedJuly 1, 2014, 3:42 p.m....Benjamin Franklin Effect, most people actually enjoy helping (just ignore the people too busy to respond to you...can do during your training is meet interesting people doing interesting things. I call this building ...insights you will get from the interaction. Most people like to help One good place to look for interesting...Sharing a school is a little connection that many people honor. Talking with strangers is usually challenging... with strangers. Be bold in asking all kinds of people to connect with you and to share their stories....uncover interesting opportunities by meeting with people working in diverse and unusual fields. My favorite... -
Easing the Protein Purification Process with pCri
TypeBlog PostPublishedJune 19, 2015, 3:08 p.m....out of protein purification through simple and efficient multiplexing. pCri plasmids are divided into two...schematic below), enabling a researcher to test multiple tags for one protein quickly and easily. These...one need to remove the tag post-purification. A sample of pCri constructs: pCri-1a, -4a and -9a. 6xHis...on these and other tags used in the pCri system, please see Plasmids 101: Protein Tags. Beyond E. coli...purification. For a more thorough introduction to pCri, please see the original publication cited below. pCri ...plasmids are available either individually or as a complete kit from Addgene. References 1. Goulas, Theodoros... -
Fluorescent Proteins 101: When GFP lets you down
TypeBlog PostPublishedAug. 23, 2018, 12:05 p.m....to the popularity of GFP including (i) fast and complete maturation to functional, fluorescent protein ...endogenous fluorescent co-factors can be used. Examples include the LOV domains that bind flavin (Buckley...under acidic conditions, with pKa values below 4. Examples include mTurquoise2 (pKa = 3.1), tagRFP (pKa =...diameter of about 2.4 nm (Hink et al., 2000). The complete beta-barrel is necessary for its fluorescence ...specifically and with high affinity to fluorophores. Examples are the FlAsH tag, the His tag and several others... bind the molecule of interest. For instance, pleckstrin homology domains can be used to detect specific... proteins are also a better choice for thicker samples. Another solution to avoid autofluorescence is ... -
CRISPR 101: Homology Directed Repair
TypeBlog PostPublishedJan. 26, 2023, 2:15 p.m....recombination intermediates can then resolve to complete the DNA repair process Homology directed repair...happen on the crossing strand (horizontally at the purple arrows) or on the non-crossing strand (vertically...is elongated and ligated to fill the gap, thus completing SDSA. Break-induced repair (BIR) pathway BIR... and even easy. This targeted-break technology coupled with HDR is a powerful site-specific knock-in tool... uses reverse transcriptase to generate the complementary ssDNA. Easi-CRISPR was shown to increasing editing...series was designed to help explain the basic principles driving CRISPRs and genome editing technology...Pfeiffer P, et al. Mutagenesis (2000). PubMed. Multiple Pathways of Recombination Induced by Double-Strand... -
CRISPR 101: Engineering the Plant Genome Using CRISPR/Cas9
TypeBlog PostPublishedOct. 11, 2016, 2:30 p.m....plants using the same general experimental design principles developed in other model organisms (see our CRISPR... CRISPR guide for common CRISPR principles). However, plant-specific modifications to commonly used CRISPR...cloning required. As with other model systems, multiple gRNAs can be expressed to modify several genomic...genomic loci at once (get more information on multiplexing gRNAs here). Cas9 is commonly tagged with a nuclear...longer RNAs (such as mRNAs for gene expression). Examples of commonly used RNA pol II promoters for Cas9...delivery and expression in planta using chitosan-complexed single-walled carbon nanotube carriers. Nat Nanotechnol...Zhang Y, Qi Y (2015) A CRISPR/Cas9 Toolbox for Multiplexed Plant Genome Editing and Transcriptional Regulation... -
Site Directed Mutagenesis by PCR
TypeBlog PostPublishedAug. 2, 2016, 2:30 p.m....are designed to be complementary, but each primer may extend beyond the complementary region as long as...with enzyme “B” detects primer multimerization (purple) which is evident by an increase in the size of... Primer Design As a rule of thumb, 11 bp of complementary sequence on either side of the desired mutation...A-overhangs (a feature used in TA-cloning). Such non-complementary bases at the 3’-end interfere with plasmid reconstitution...that any residual parental plasmid (usually from incomplete DpnI digest) can also form colonies under these...particularly relevant in cases where a large plasmid with multiple functional regions is used as template for the.... Breslauer, Kenneth J., et al. "Predicting DNA duplex stability from the base sequence." Proceedings ... -
An Addgene Co-op: The Intersection of Fun, Friendship, and Work
TypeBlog PostPublishedJune 29, 2023, 7 p.m....found the people at Addgene have become my friends, mentors, and some of my favorite people to talk to...student at Northeastern University, I am required to complete a six month work experience called a co-op to ...work of others. Having an environment filled with people who are interested in understanding the work you...Thankfully, my managers set aside time for me to complete an online social media certification on social...changes to our social media strategy. We ended up implementing some of these changes and seeing early results...results that suggest our content is reaching more people who need it! After a few months, we wanted to share... -
New Viral Vectors - Fall 2024
TypeBlog PostPublishedOct. 3, 2024, 1:15 p.m....Optogenetics Ofer Yizhar New viral prep with multiple serotypes pAAV-Syn-FLEX-rc[ChrimsonR-tdTomato...Biosensors GENIE Project New viral prep with multiple serotypes pGP-AAV-GFAP-iGABASnFR2(no bind)...Biosensors GENIE Project New viral prep with multiple serotypes pGP-AAV-syn-iGABASnFR2-WPRE AAV1...Biosensors GENIE Project New viral prep with multiple serotypes pGP-AAV-syn-iGABASnFR2(no bind)-...Biosensors GENIE Project New viral prep with multiple serotypes pGP-AAV-syn-flex-iGABASnFR2-WPRE...Biosensors GENIE Project New viral prep with multiple serotypes Looking for more viral vectors... Multiple Authors... -
Build Your CRISPR Vocabulary
TypeBlog PostPublishedJuly 2, 2024, 1:15 p.m.... forming the repeat:antirepeat duplex. tracrRNAs also contain multiple stem loops that are recognized ...Palindromic Repeats. That contrast of simplicity and complexity is reflected in the biology, too. CRISPR is an..., and the spacer sequence is used to target complementary DNA sequences. In laboratory settings, an sgRNA...whose sequence is not checked. Assuming full complementarity, its sequence matches the guide sequence. PAM...either crRNA or sgRNA). Cas9 requires perfect complementarity in this region between the guide and target...post on the structural biology of Cas9. Want a simpler guide to keep at your desk? We’ve got a CRISPR ... -
Oh, The Places You Can Go: Careers in Science Communication - Science Writing
TypeBlog PostPublishedOct. 3, 2017, 12:11 p.m....defining a couple of key terms is a better approach. Also, appealing images and colors draw people in. Drawing...webinars—I have them stacked close together for the next couple of months. This involves a lot of communication...of science communications-- Be able to talk with people: “If you do this for a living, science writing ... when I am in new territory. Perhaps there are examples of science writers who can get everything that...non-scientists: “Jargon is a killer for science literacy. If people have to understand a lot of big words to access...mechanism that also looks nice is a great way to bring people in.” Are you interested in careers outside academia... -
Advice for Choosing a Research Project
TypeBlog PostPublishedApril 17, 2014, 2:45 p.m....research project can take several years to complete and has multiple benchmarks. Think about what your personal...professional stage of your career by connecting you with people of similar interests and goals. The following questions...field? You want to choose a research question that people (especially you!) care about. Have you critically...reach these milestones? Can you get assistance from people who have the required expertise to help move your... of keeping up-to-date with the literature of a couple of well-known journals that discuss general science...Researchers should pick a topic that “looks at the complexity of biology and break apart a piece that can address...