...industry and how delegating can empower people Chapter 3: How Crystal advanced at Novartis, and how to prepare...Intro Chapter 1: Who is Crystal Shih? Crystal's educational background and position at Novartis. Chapter ...Chapter 7: How Crystal developed her science communication skills and how they've helped her at Novartis...

...ensure nothing is covering it. Turning on the UV light 3-4 times a week for up to half an hour can help keep...Journal of applied phycology27.6 (2015): 2321-2332. 3. Contamination, Ileana S., John P. S., Basic Methods...results. Most cell culture media contain the pH indicator phenol red (pH range 6.0 (yellow) - 8.0 (red))...Anything between orange and light orange usually indicates that the cells have grown enough, are making their...

...adenovirus genes which help mediate AAV viral replication; and 3) the transfer plasmid which encodes a transgene...ITR-containing plasmid. This whole process takes ~3 hours to complete: 1 hours hands-on time and 2 hours...which provides the AAV replication (rep) and capsid (cap) genes. AAV replication uses the host’s polymerase...this AAV purification video! It didn’t exist when I did my first iodixanol gradient purification, but I ...In this post, I’ll share some AAV production, purification, and titration tips, while also summarizing ... that aids in the homologous recombination of duplicated regions like the ITRs. Restriction digest screening...consider performing PEG precipitation. Instead of sonicating the cell pellet, I lyse the cells with this AAV...

...Professional Certification Institute (GPCI). GPCI evaluates a candidate's expertise (a minimum of 3 years experience...the organization(s) you serve time and resources. 3. Get a mentor, find your people Find someone who is...development, certification, degrees, foundations, coaching and even Twitter Chats dedicated to the Grant...the candidate application process, the candidate’s ability to write a grant application through a written... much more than writing. In fact, many grant applications and grant makers have moved away from long written...Now they require grant seekers to use online applications with significant character limits for the written...written portion of the grant application. They have increased their demand for stronger budgets with clear...

... enhancers.  3. Measuring temporal changes in chromatin interactions A third application for CAPTURE 2.0...expressing 1) FLAG-biotin-tagged dCas9, 2) BirA, and 3) one or more gRNAs Two separate lentiviruses delivering... gene’s transcription. Sometimes enhancers are located thousands of base pairs away from the gene they...created a dCas9-based CAPTURE (CRISPR Affinity Purification in situ of Regulatory Elements) method. The ...could only detect chromatin interactions at one location in the genome at a time (Liu et al., 2017). Recently...at once (Liu et al., 2020). CRISPR Affinity Purification in situ of Regulatory Elements: The original...biotin ligase a gRNA(s) targeting a single genomic location of interest  When these three components are ...

...prioritize investments over the next 3, 5, or 10 years. Complicating these challenges is the fact that they... Consulting firms deploy case teams, usually with 3-6 consultants, who work on analyzing the situation...advancements. Consultants can help these teams develop communication strategies, or understand, and publish on the...success. Or the client might need help with the intricate task of designing a strategic vision for a portfolio...chemotherapies, or developing innovative mobile phone applications and services that help diabetic patients manage...knowledge from previous consulting experiences, educational training, and conferences, and synthesizing this...that gives the client a defined path forward. Communication is key as ideas must be relayed quickly both...

...had therefore been trafficked between locations). Figure 3: A) LOV-Turbo becomes active when illuminated... h, and PMA activation induces cFos::GFP-iPAK4 at 3 or 6 h. Scale bar 5 µm. Images adapted from Lin et...Figure 1: Choosing the best PE6 variant for your application. Image reused form Doman and Pandey et al., 2023... CreVLPs, and eVLPs for in vivo and in vitro applications.   Figure 2: Modular features of DIRECTED...approach for experimental therapies and medical applications.  Find DIRECTED plasmids here! Strebinger, D...delivery by modular envelope design. Nature Communications, 14(1), 5141. https://doi.org/10.1038/s41467...interactions or map a local proteome, but some applications have been challenging or inaccessible. The Ting...

...enhancers 3. CRISPR out the enhancers from step 2. See which ESCs die or differentiate Step 3 reveals which...helps us to locate interesting addresses in the genome. Let’s now examine how we can locate powerful stretches..., start, and stop. For instance (chr3, 1, 10) indicates a stretch of DNA at the very beginning of the ...for the genome" techniques Technique What It Locates ChIP-seq Proteins (for our example later, histones...therapeutics. We’ll have more to say about clinical applications of CRISPR in the next section. In order to denote...’s associated histone proteins K4 and K27 – locations of the amino acid lysine in H3 me1, me3, and...aceylation, respectively (these are chemical modifications, or functional groups, found on lysine) Putting...

...propagation along dendrites for the first time (Figure 3). The team also used AAV9 to deliver ASAP5 to the ...microscopy at depths up to 600 µm. Figure 3: ASAP5 for voltage imaging. A) ASAP5 selected for ...single-molecule biophysics Fast and responsive voltage indicator ONE-GO biosensors kit Antibodies for neurodegeneration... an enhanced pU6cm promoter, viral replicon amplification of cargo, and heat treatment.  Figure...2024.12.20.629623.  A fast and responsive voltage indicator with enhanced sensitivity for unitary synaptic...and more responsive genetically-encoded voltage indicator (GEVI) than previous versions. ASAP5 can be imaged...The initiation site identified in panel (C) is indicated; red line is exponential fit. Adapted with permission...

...31668202421', style='text-align: right;' %} Figure 3. An animated map showing the plasmid orders and new...were zero plasmid requests. Afterwards, from 2/24/20–3/15/20 (the supposed end of lockdown), request activity...subfields.  Plasmid requests correlate with paper publication The dataset includes 11,426 plasmid requests ...14%). Spikes in plasmid requests correspond to publications of popular papers. Figure 1: Plasmid requests...effect on plasmid requests Before we began any sophisticated analyses, we first visualized our data in various...orders and new COVID-19 cases for each week by location. We observed that as expected, plasmid orders ...independence assumption of the Poisson process. This modification also eases the interpretation of the parameters...

...PMID 25282355. PubMed Central: PMCID: PMC4317352. 3. Beisel, Chase L., Ahmed A. Gomaa, and Rodolphe Barrangou...Targeted Bacterial Population Editing.” Cell Syst. 1(3):187-196. (2015). PubMed: PMID 26973885. Additional...containing a phage origin of replication that can be packaged into replication-incompetent phage particles...essential processes: cell wall synthesis, DNA replication, or protein synthesis. Because they target such... and medical organizations are working to help educate the public about proper antibiotic usage. Although...mammalian viral vectors in that the vectors are replication-deficient. Thus, a very large number of phagemids...CRISPR antimicrobials combines them with both replication-competent phages and antibiotics. Phage therapy...

...sequence only functions at the extreme C-terminus [3]. You must therefore determine your cloning strategy...PMID: 19819147. Pubmed Central PMCID: PMC2784028. 3. Munro, Sean, and H. R. Pelham. "Use of peptide tagging...compartments undergo significant post-translational modifications including glycosylation, disulfide bond formation...are equally susceptible to these non-native modifications, which can all potentially affect function [...N-glycans [9]. Both of these posttranslational modifications can disrupt FP folding rendering the protein... non-optimized parental proteins. Our recent publication illustrated the quantitative increase in fluorescent...and Jack Griffith, her research focuses on the replication machinery of Kaposi’s sarcoma-associated herpesvirus...

...and makes your experimental setup less cumbersome. 3. Portability across multiple systems Terskikh et al...PMID: 19889833. PubMed Central PMCID: PMC2801723. 3. Verkhusha, Vladislav V., et al. "An enhanced mutant... (green or blue) within a population of timer indicates the age of the pool: the higher the ratio, the...-FT-N1 pTRE-Slow-FT pBAD/HisB-Slow-FT -  Applications of fluorescent protein timers FP timers have...biological event (trafficking to a subcellular location, start of gene expression, development of a cell... insulin-secreting beta-like cells." Nature communications 7 (2016). PubMed PMID: 27063289. PubMed Central...Learn More About Fluorescent Protein Aggregation Catch Up On Your GFP Basics Get Advice on Choosing Your...

...interfere with it in a BSL-2 lab instead of a BSL-3 lab. Jesse Bloom’s lab published a protocol and reagents... CRISPR-Based Detection Tools. The CRISPR Journal 3:142–145. https://doi.org/10.1089/crispr.2020.0030 ...a method that uses loop-mediated isothermal amplification after reverse transcription (RT-LAMP) and a ...receptor for SARS-CoV. The lab also found that cathepsins play a role in SARS-CoV infection and deposited...deposited plasmids for the expression of cathepsin S, L, and B. This became a great starting point for SARS-CoV..., Zengel JR, Sathyan V, Nguyenla X, Van Dis E, Catamura C, Yamashiro LH,  Begeman A, Stark JC, Shon DJ...

...the gel and add gel components to polymerize. Step 3: digest the sample to allow for even expansion.  ...:832–863. https://doi.org/10.1038/s41596-018-0117-3 Wassie AT, Zhao Y, Boyden ES (2019) Expansion microscopy...with an abundance of protocols and examples of applications from across disciplines now available. In this...experiment or evaluating ExM images in a new publication.         Why Expansion Microscopy?  Traditional...microscopes sounds great, right? But there is always a catch - some limitation of the technique that users should...Tissue expansion is not compatible with in vivo applications and there’s just not much to be done about that...

...pathway (Charpentier et al., Nat Com).     Fig. 3: HDR events compete with NHEJ-mediated edits of small...Non-Homologous End Joining Learn about Base Editing Read 3 Tips to Improve HDR Efficiency for CRISPR Editing ...cut site location Position your CRISPR cut site as close as possible to the knock-in location. HDR depends...sequence to be incorporated at a precise genomic location, with little to no room for error. It’s no surprise...nutrients in their media, they will not undergo DNA replication, so make sure you are keeping an eye on those...

...assessment of many protein targets in a single sample; 3) producing recombinant, partially humanized rabbit...target proteins of interest for labeling, quantification, purification, chromatin immunoprecipitation and more...created the Monoclonal Antibody Research Institute dedicated to: 1) developing new ways of generating antibodies...Freedom: The Expansion of Data Sharing in Research Publications ...

...into Stage 3 or Stage 4 cancer. These later stages of a cancer diagnosis indicate, for Stage 3, that the...large in size and may have spread while Stage 4 indicates that the cancer has spread from where it originated... dire prognosis for mesothelioma patients is to catch the disease earlier. New techniques are currently... working to fight the cancer.  Beyond early modifications to standard treatments and early detection, ...

... for an extended period of time (>10 passages or >3 months). Other times cell lines are unavailable from...State University of New York Press, Albany (1986). 3. Masters, John R. "HeLa cells 50 years on: the good...Marker-independent identification of glioma-initiating cells. The International Cell Line Authentication Committee...propagation of the line, and can include assays for authentication and contamination) and cell lines can become...let alone the human cost. Here are just a few publications which have since been retracted due to cell ...

... kb at most - which means that a gene larger than 3 kb cannot be delivered using this vector. For example...emerges from disgrace to be the next big thing, again 3. Acland G. et al., 2001. Gene therapy restores vision... time advances in human genetics led to the identification of several disease-causing genes, including... But it is the hard work and persistence of a dedicated team of scientists led by Dr. Jean Bennett (UPenn...-1250015778 7. Yu W and Wu Z., 2018. In vivo Applications of CRISPR-Based Genome Editing in the Retina...