...." F1000Research 3 (2014). Pubmed PMID: 25653845. Pubmed Central PMCID: PMC4304227. 3. Sauermann, Henry...positions within 5-7 years of earning their PhDs (3), showing that the ‘standard’ pathway into academia..., Eve. "Looking out for future scientists." eLife 3 (2014): e04901. Pubmed PMID: 25291257. Pubmed Central...University of Massachusetts Medical School. Graduate education programs are also beginning to address these issues...The panel discussion on Friday morning will be dedicated to Academic data and the Labor Market. On this...Sinche is a STEM career counselor, Director of Education for the Jackson Laboratory for Genomic Medicine...disease.  Outside the lab, he is interested in communicating science and in the intersection between science...

...three components (Figure 1): 1) Exo, 2) Beta, and 3) Gam. All three are required for recombineering with...DNA introduced into the E. coli. Exo: Exo is a 5’→3’ dsDNA-dependent exonuclease.  Exo will degrade linear... 1) a partially dsDNA duplex with single-stranded 3’ overhangs or 2) if the dsDNA was short enough, a ...the translated protein.  These changes can be 5’ or 3’ of the desired change.   Table 1: Overview of ds-... integrated defective prophage 2) from a plasmid, 3) from mini-λ or 4) from the lambda red phage itself...PubMed PMID: 10811905.PubMed Central PMCID: PMC18544. 3. Murphy, K. 2016. λ Recombination and Recombineering...of its biggest limitations is that sequence modifications can only be made at restriction enzyme cut sites...

...crRNACpf1 Endonuclease size spCas9: ~4 kbsaCas9: ~3 kb ~3.9kb crRNA/gRNA length gRNA: ~100 nt crNA:... Lastly, an AAV vector was used to multiplex edit 3-4 genes in a primary culture of mouse neurons and ...and GFP.  Of these neurons, ~15% had indels at all 3 targeted loci.  See the chart below for a summary ...Endonuclease of a Class 2 CRISPR-Cas System. Cell,163(3), 759-771. PubMed PMID: 26422227. PubMed Central PMCID... a simple and effective method with multiple applications (in vitro, in vivo, transfection, viral transduction...Resources Find CRISPR Plasmids for Your Research Catch up on Your CRISPR Background with Our Guide Pages...

...where the lab magic is happening. 3) Volunteer – Step into science education Do you aim to reach young people...-to-day business with popular work experiences of 3-10 days. And the Society of Experimental Biology offers...science communication are as varied as the field itself. Among other things, science communication comprises...certain aspects of art and education. If you suspect that science communication might be for you, don’t ... networking with your local education community. Get involved in education advocacy non-profits like Phipps... Training – Get a formal education Some people get into science communication by taking a media or journalism...project. Science communication opens doors Getting involved in science communication has been the most...

...PMID: 27175853. Pubmed Central PMCID: PMC4866822. 3. McDowell, Gary S., et al. "Shaping the Future of ...perspective from junior scientists."F1000Research 3 (2014). Pubmed PMID: 25653845. Pubmed Central PMCID...Riglar Advocating for Science Symposium and Workshop 2016 – Sept 16-17 MIT, Boston The Advocating for Science...futureofresearch.org/advocating-for-science-boston-2016/. Sign Up For the Advocating for Science Symposium...Director of Media Communications, Harvard University Ray Howell -Founder of Howell Communications Philip Brenner...union dues. Time will tell the extent of the implications of this ruling and whether graduate students...labor laws, rather than as trainees in a special category, as has been the ‘labor-of-love’ philosophy of...

...field, (2) that you have three 3 years of research experience in the field, (3) that you have an offer of ...employment based green card applications and approved 91,023 during the first 3 quarters of 2017. As an international...job offer is needed for this category. To qualify, you need to satisfy 3 out of the 10 criteria listed...well positioned to advance the proposed endeavor; (3) The foreign national’s proposed endeavor will be ...institute. For the EB-1B, you need to prove 3 threshold requirements: (1) that you have international ... you must show that your company employs at least 3 full-time people engaged in researcher activities....higher education nonprofit entities related to or affiliated with institutions of higher education nonprofit...

...MSOE center for BioMolecular Modeling and the CEO of 3-D Molecular Designs. Have you ever held your favorite...models were useful in that they not only traced the 3-D path of the protein’s backbone in three-dimensional...arginine 53 really is right there, at the end of helix 3…just like we said it was”.   Seeing is believing. ...MSOE center for BioMolecular Modeling and the CEO of 3-D Molecular Designs.     Additional Resources on the...development programs for bioscience educators. Using these hands-on tools, educators introduce their students to...biologists have always struggled to effectively communicate the nuances of protein structure in a flat 2-...dangerous. Subscribe to CRISPR Blog Posts How do educators and students use molecular models?   We created...

...plasmids ready for deep mutational scanning.     3 reasons to be interested in deep mutational scanning...polymerase, 2) increased PCR extension time, 3) increased concentration of MgCl2, 4) increased concentration ...synthesize a complementary mutant strand (step 2 and 3). Then the uracil-labeled template strand is degraded...mutagenesis method are 1) ease of use, 2) time savings, and 3) ability to perform single site or multi-site saturation...Mutagenesis Protocols, pg. 103-109. PubMed PMID: 20676978. 3. Wrenbeck, Emily E., Justin R. Klesmith, James A. ...created by making some or all of the following modifications: 1) increased concentration of Taq polymerase...

... seconds for 3 hours. Image from Lee et al., 2020. SPOTlight was used to image over 3 million cell...investigated three Casɸ orthologs, Casɸ-1, Casɸ-2, and Casɸ-3. This system is notable for several reasons. Casɸ ...For instance, for Casɸ-2 the PAM sequence is 5’-TBN-3’ (where B is G, T, or C). The authors demonstrate ...including excision or insertion, inversion, translocation or cassette exchange. By utilizing recombinases...BioRender.com In their recent ACS Synthetic Biology publication, Mary Dunlop’s lab generated a photosensitive...regions with Vivid (Vvd) photodimers. Upon the application of blue light, Vvd dimerizes bringing the C-terminal...

...mutations in RPE65 (n=3), showed significant improvements in vision over a period of 1 to 3 years – the treatment...neutralizing antibodies against some AAV types, AAV1, 2, 3 and 5 are thought to be as high as 70% among the human...PMID: 20463753. PubMed Central PMCID: PMC3152950. 3. Jacobson, Samuel G., et al. "Improvement and decline...passionate about HIV cure research and science communication.     References 1. Gaspar, H. B., S. Howe, and...affinity-purified receptor-targeted viral vectors." Nature communications 6 (2015): ncomms7246. PubMed PMID: 25665714...our AAV Production Protocol Check out our AAV Purification Protocol ...

...yeast as an alternative source of essential omega-3 fatty acids complementing vegetable oil, a somewhat...PMID: 27984002. 2. Bilan, Dmitry S. et al. “HyPer-3: A Genetically Encoded H2O2 Probe with Improved Performance...Lifetime Imaging” ACS Chemical Biology 2013 Mar 15;8(3):535-42. PubMed PMID: 23256573. Additional Resources...scientists work on projects spanning from the identification of plant compounds and insecticidal proteins...bacteria. Learn more about the different CRISPR applications in plants.     Transgenic yeast as source of... yeast capable of desaturating fatty acids. Identification of Insecticidal Proteins Linh Bao Ton (M. Sc...Arturo Rivera Solís, Universidad Autónoma de Yucatán, Mexico Dr. Rodrigo Arturo Rivera Solís, Associate...

...in plants. When expressed, the construct produces 3 enzymes from a single mRNA using 2a sites. These convert...Betacyanin is produced from tyrosine by the activity of 3 enzymes and a spontaneous step. (B) RUBY is a reporter...collect seed for further experiments.     Fig. 3: (Top right) A RUBY-expressing Arabidopsis seed following...markers for plant transformation fall into two main categories: antibiotic/herbicide resistance and fluorescent...culture), which is great for a wide variety of applications. But some techniques require you to select against...these mutant plants. Using RUBY to simplify the identification of these mutants will speed up my work and ...student studying plant development and cell-cell communication in the Nimchuk Lab in the Biology Department...

... and breathing, and often results in death within 3-5 years from symptom onset. Disease mechanisms are...loci seem to be associated with the disease and implicate a wide range of cellular processes. The vast majority... consists mainly of managing symptoms through medication and surgery. PD primarily involves the malfunction...available from Addgene, sortable by type of genetic modification and target species. You can also find our guide...Sun et al. Nat Methods. 2020 Oct 21. Glutamate indicators with improved activation kinetics and localization...A multicolor atlas for whole-brain neuronal identification in C. elegans. Yemini et al. Cell. 2021 Jan...window) A news website and information resource dedicated to helping researchers accelerate discovery and...

...deciding which one to buy. Table 3: Comparison of popular plasmid purification kit options Manufacturer...it easier to compare kits at a quick glance, Table 3 below contains details regarding kits from a variety...of some commonly used kits, please note that Table 3 is certainly not comprehensive — there are many other...column-based DNA purification methods perform well in terms of quality and yield. Bead-based purifications tend to...standard purification kits, there’s a range of specialty kits available for plasmid purification. Kits can...optimized for purification of these large plasmids, while others have suggested modifications to their normal...you’re new to using purification kits, or if you’re struggling with a purification, reading the full instruction...

...imaged the mouse brains, they found many more than 3-4 colors due to the tandem integration of multiple...colorful technology. Optimizing the system: Brainbow-3 Although Brainbow provided neuroscientists with the...Looger LL, Simpson JH. Nature Methods. 2011 Mar;8(3):253-9. doi: 10.1038/nmeth.1566. Epub 2011 Feb 6. ...Dickson BJ, Salecker I. Nature Methods. 2011 Mar;8(3):260-6. doi: 10.1038/nmeth.1567. Epub 2011 Feb 6. ...Beaurepaire E, Morin X, Livet J. Neuron. 2014 Feb 5;81(3):505-20. PubMed.   ...of farnesylated derivatives enables labeling of delicate axonal and dendritic processes not previously ...Between Brainbow Systems   Brainbow variants and applications beyond the mouse brain In addition to improving...

...DNA primers that bind upstream (5’) and downstream (3’) of the DNA region of interest. Primers are crucial... the dNTPs.  Let’s jump into the specifics of the 3 basic steps of a PCR reaction:  Denaturation- To...extension is repeated 25-35 times to exponentially replicate the target DNA of interest. This entire reaction...have been developed for different scientific applications. These PCR methods all use the same basic PCR...the final cycles which would slow or stop PCR amplification. Other times, the PCR reaction may not be 100%...quantified by qPCR. qPCR is used in a variety of applications including gene expression profiling, studying...concentration, unlike qPCR where results can vary across replicates. ddPCR can be used to quantify DNA sequences ...

...organs. Restraint: scruffing Location: peritoneal cavity Gauge: 25–27 Volume: < 2–3 ml Skill required: intermediate...or mechanical restraint Location: back or hind legs Gauge: 25–27 Volume: < 2–3 ml (~1 ml per site) Skill...Intraperitoneal Scruffing Peritoneal cavity 25–27 < 2–3 mL Intermediate Intramuscular Mechanical restrainer...Back or hind legs; between skin and muscle 25–27 < 2–3 mL (~1 mL per site) Beginner Intradermal Anesthesia.... It sounds complicated but has a pretty low learning curve! Restraint: scruffing Location: throat Gauge... Restraint: mechanical restraint or anesthesia Location: tail vein or retro-orbitally Gauge: 27–30 Volume...practice to ensure the proper injection site has been located and to avoid potential off-target pokes to nearby...

...we look closely at Figure 3, which shows requested CRISPR plasmids categorized by “age” or year they were...    Figure 3: Percentage of requests per year for CRISPR plasmids categorized by year of deposit...2, and the longevity of tools seen in Figure 3, indicates that the usefulness of popular CRISPR tools ...plasmids requested each year by year of deposit (Figure 3), we saw that once CRISPR plasmids become popular,...estimated scatterplot smoothing. (C) Number of requests per year for pooled libraries, categorized by function...insertion via a chemical modifier. We used these categories to see what kinds of CRISPR tools people were... pooled libraries, although we used different categories (Figure 2C). Note that in Figure 2, the numbers...

...sequence (Zong et al., 2018) (see CBE-3 in Figure 3). Figure 3: Activity windows of plant base editors...position 4-8 counting the ‘NGG’ PAM as 21-23 (Figure 3). If the target ‘A’ is closer to the PAM, i.e., positioned... 353.6305 (2016): aaf8729. PubMed PMID: 27492474. 3. Gaudelli, Nicole M., et al. "Programmable base editing...          The artificially evolved adenosine deaminase catalyzes the conversion of a target ‘A’ to ‘I’ (inosine... optimize base editing efficiency based on the location of the ‘A’ relative to the protospacer adjacent...Choose a base editing platform depending on the location of your target base in the protospacer sequence...

... point for a new generation of ABEs), consists of 3 components:  a deaminase (TadA, originally from E.coli...al., 2020). The generated ABE8s needed to produce 3 concurrent base edits to withstand antibiotic selection...protospacer adjacent motif (PAM) made up of 5′-NGG-3′ (“N” can be any nucleotide base, followed by 2 guanines...ABE8.20-m was the winner. It could simultaneously edit 3 genes (98.1%, 98.3%, or 98.6% efficiency, respectively... E.coli, named TadA7.10 in ABE7.10)  a catalytically impaired Cas protein (dCas or Cas nickase)  a guide...accidental insertions and deletions (indel) by the catalytically impaired D10A nickase from ABE7.10 can be a ...the authors created ABE8 constructs using a catalytically “dead” S. pyogenes Cas9 (dC9-ABE8.x-m/d). The...