We narrowed to 788 results for: lat
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TypeBlog PostPublishedNov. 18, 2014, 7:15 p.m....sessions, exhibit booths, happy hour socials, and late, late night socials. As a newbie to the SfN conference... global coffee supply… My back-of-the-napkin calculations put the daily consumption here at ~ 60,000 cups...students and postdocs, which accounts for many, many lattes and espressos. The energy boost is well needed ... Investigator Awards and we couldn’t wait to congratulate him on his successes. We've been lucky enough...SfN14 on Twitter for live tweeting updates and the latest news from attendees. And for those of you presenting...
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A Scientist's Guide to Ebola
TypeBlog PostPublishedOct. 16, 2014, 3:01 p.m....have similarly curated their Ebola-related content. -And the latest updates on the ongoing outbreak from... underestimate. Given all the excitement and speculation, we thought we’d do a little digging and assemble...assemble a rough guide to the science of Ebola and related resources. Ebola Basics A few basics: Ebolavirus... data on five strains of Ebola representing the latest outbreak and earlier ones, along with links to ...the current Ebola crisis, Science and Science Translational Medicine have made their complete collection... -
New Optogenetic Tools for Cytoskeleton and Membrane Control
TypeBlog PostPublishedSept. 28, 2023, 1:15 p.m.... target to specific subcellular regions, laser ablation can damage other cellular structures, and genetic...genetic mutations are not easily reversible. Late last year, the Akhmanova Lab published a new optogenetic...for a wide variety of experiments. Opto-RhoGEFs manipulate cell morphology and adhesion But what if you’... Taking control of a RhoGEF is a great way to stimulate and study these physical processes. Joachim Goedhart...of membrane-recruitment optogenetic tools for manipulating Rho GTPases based on BcLOV4. These single-component...membranes) (Figure 4). The authors used SuperPLD to manipulate phosphatidic acid signaling (such as Hippo, AMPK...minutes of blue light illumination, substantial PA accumulates in the plasma membranes, indicated by the PA ... -
CRISPR 101: Engineering the Plant Genome Using CRISPR/Cas9
TypeBlog PostPublishedOct. 11, 2016, 2:30 p.m....often requiring several rounds of selection to isolate plants with the phenotype of interest. Genome engineering... modification of known or suspected genes that regulate a desired phenotype. In fact, CRISPR has already...variants have been created in an effort to increase translation in a particular plant species or cell type (Belhaj...driven by plant-derived RNA pol II promoters which regulate expression of longer RNAs (such as mRNAs for gene...plant cytorhabdovirus as versatile expression platforms for planthopper and cereal genomic studies. New...Multiplexed Plant Genome Editing and Transcriptional Regulation. Plant Physiol 169:971–985 . https://doi.org/...(2016) Gene-edited CRISPR mushroom escapes US regulation. Nature 532:293–293 . https://doi.org/10.1038... -
Evolution of Brainbow: Using Cre-lox for Multicolor Labeling of Neurons
TypeBlog PostPublishedApril 24, 2015, 2:39 p.m.... be the hot new way to manipulate gene expression, but other gene manipulation systems remain valuable...To achieve even cell labeling, they generated farnesylated derivatives of the fluorescent proteins, which...trafficked to cell membranes. Membrane trafficking of farnesylated derivatives enables labeling of delicate axonal... default fluorescence due to the addition of translation-blocking STOP cassette immediately following ...woodchuck hepatitis virus post-transcriptional regulatory element (WPRE), commonly used to increase transgene...and Flpbow can be used to label distinct cell populations. To decrease the animal breeding necessary to... -
Optogenetics + CRISPR, Using Light to Control Genome Editing
TypeBlog PostPublishedSept. 3, 2020, 12:15 p.m....formed 20-40 min later. The CASANOVA system is versatile in that it can be adapted to regulate gene editing...modifications by NHEJ and HDR using split systems Later in 2015, the Sato lab unveiled a photoactivatable...opens intriguing possibilities for optogenetic manipulation of CRISPR-based systems in animal models. Use... prevent DNA binding in the dark. Upon light stimulation, the pdDronpa moieties move apart, allowing DNA...LOV2 has a C-terminal helix that unfolds when stimulated by blue light. When fused to the catalytic site...genome and chromatin architecture, it is useful to regulate the time in which dCas9 binding occurs. Bubeck...Niopek D (2020) Optogenetics and CRISPR: A New Relationship Built to Last. In: Methods in Molecular Biology... -
Scientist Networking: What is an Informational Interview?
TypeBlog PostPublishedJuly 1, 2014, 3:42 p.m.... more open to developing a relationship. The time to develop relationships is before you need them (well...doing interesting things. I call this building relationships because networking has gotten a bad reputation...Scientists enjoy learning new things. Building new relationships is all about learning new things from other ...networking for a job connection with ongoing relationship building through Informational Interviews. Making...is always best to meet in person to develop relationships, but if it is not possible consider a phone ... -
Easing the Protein Purification Process with pCri
TypeBlog PostPublishedJune 19, 2015, 3:08 p.m....has the capacity to glycosylate proteins and may recapitulate the post-translational modifications added...requires a substantial amount of properly folded, relatively pure protein, but getting to this stage is often...one’s protein of interest can save a lot of time later. Protein purification specialists recommend doing...the MISTIC (membrane-integrating sequence for translation of integral-membrane protein constructs) protein... -
CRISPR Protocol for Genomic Deletions in Mammalian Cell Lines [Video]
TypeBlog PostPublishedFeb. 18, 2015, 3:09 p.m....non-coding regulatory elements. Regulatory DNA function is typically inferred based on correlative biochemical...of the method. One observation was an inverse relationship between deletion size and frequency [3]. There...interested in exploring the function of non-coding regulatory elements in their native chromatin context. An... Like the video? The Orkin and Bauer lab's related paper describes their methods for CRISPR design... -
Scientific Sharing in the Time of COVID-19: Databases and Resources
TypeBlog PostPublishedMay 12, 2020, 1:15 p.m....asked the individuals behind some of these new platforms what their motivations were for developing these...knowledge that could help fight COVID-19. They’ve also collated a list of useful research resources. "We realized...Canadian volunteers, and features over 200 COVID-19 related research projects, funding opportunities, and community...community response initiatives. Accelerate the translational impact of research Universities have also been...creating research synergies and accelerating the translational impact of research activities. By streamlining...research in their field including datasets and platforms for discussing new developments, and more. As ... -
CUT&RUN: An Improved Method for Studying Protein-DNA Interactions
TypeBlog PostPublishedFeb. 13, 2018, 2:51 p.m....organisms and cell lines. Although ChIP-seq is a relatively simple and robust protocol it does have limitations...study how differential access to transcriptional regulatory elements is controlled during development using...fused micrococcal nuclease (MNase) to specifically isolate DNA that is bound by a protein of interest. First...First, nuclei from tissue or cell culture are isolated using lectin-coated magnetic beads. Nuclei are then...MNase requires for its nuclease activity and is chelated from the reaction up until this point. The nuclease...simple lab protocols Browse CRISPR plasmids for isolating a genomic locus Browse FRET plasmids for studying... -
Arabidopsis in Education: How the Arabidopsis Biological Resource Center Brings Plant Science to Life
TypeBlog PostPublishedApril 12, 2018, 1:02 p.m....seeds, DNA and other resources for Arabidopsis and related species. Located at The Ohio State University in...member of the Brassicaceae family, Arabidopsis is related to a variety of food crops such as broccoli, cabbage...utility to science, and its well-known edible relatives, it is fair to say that most people do not recognize...teaching community through the TRAINED initiative (Translating Research on Arabidopsis Into a Network of Educational...through TAIR with a new ABRC database and ordering platform soon to be released. For individuals interested...Arabidopsis is small, has a short life cycle, and is relatively easy to grow, it is an ideal organism for teachers... -
Plasmids 101: Origin of Replication
TypeBlog PostPublishedNov. 11, 2020, 2:23 p.m....balance between positive and negative regulation and can be manipulated with mutations in the replicon. For...produce just a few copies depending on how they are regulated. Generally, control of replication is referred...stringent" depending on whether the ori is positively regulated by RNA or proteins, respectively. A plasmid's ...plasmid copy number Although the sequence and regulation of the ori dramatically affect the copy number...SB, Furman JL, Xiao H, Schultz PG. A versatile platform for single- and multiple-unnatural amino acid ... -
Viral Vectors 101: Parts of the AAV Transfer Plasmid
TypeBlog PostPublishedAug. 11, 2020, 1:15 p.m....polyA The polyadenylation or polyA signal aids in the nuclear export of RNA and RNA translation, and promotes...plasmid. Parts shown in grey are optional cis-regulatory elements. Key elements of an AAV plasmid...plasmid harbors non-coding sequences. Other cis-regulatory elements Next let’s talk about some of the elements...Woodchuck Hepatitis Virus (WHP) Posttranscriptional Regulatory Element (WPRE) forms a tertiary structure when...Technologies and Methods for Targeted Neuronal Manipulation. Front Neuroanat 13: . https://doi.org/10.3389... -
Hot Plasmids and Viral Preps - September 2021
TypeBlog PostPublishedSept. 21, 2021, 1:15 p.m....viral preps Improved cell-free transcription/translation system to examine biosynthetic pathways By Gabrielle...Gabrielle Clouse Recently, cell-free transcription/translation systems (also referred to as TX-TL systems) have...In summary, this cell free system allows for a relatively simple, controlled “one-pot” reaction capable...plasmids that helps identify base edited cell populations without end-point sequencing assays. These plasmids..., or XMAS-TREE, to identify base edited cell populations without end-point sequencing assays. This tool...different vectors and flow cytometry is used to sort populations by mCherry and GFP expression to check for ABE... -
Plasmid Cloning by PCR
TypeBlog PostPublishedMarch 29, 2016, 2:30 p.m....DNA, a cDNA library, or by RT-PCR. Isolate the PCR product Isolate your PCR product from the rest of the...function in the same buffer, as this will save time later In our example, we will use EcoRI and NotI to ...TGGCATATCTCGAAGTACTGAGCGGCCGCTAAGCA-3’) into this calculator we get a final Reverse Primer sequence of 5’-...If you are amplifying from a plasmid or simple template, there is very little chance for mis-priming, ...now ready for restriction digestion. As such, the later steps in this process are the same as those discussed... -
Quick Guide to All Things Lentivirus
TypeBlog PostPublishedMarch 21, 2017, 2:30 p.m....of interest as well as sequences necessary to encapsulate that insert into a viral particle (the packaging...lentiviral vector system is the tetracycline (Tet)-regulated system. You can choose either the Tet-off system...rtTA (FUW-M2rtTA). In this kind of approach, a population of cells might express only the tTA or your transgene...proto-oncogenes or oncogenes for instance should be manipulated with care. See the NIH’s biosafety guidelines...viral capsid antigen p24, the FACS dosing method correlates viral titre to the number of cells transduced...order to aid viral binding. This method, called spinoculation, has been found very effective for T cells transduction...Yasutsugu Suzuki and Youichi Suzuki (2011). Gene Regulatable Lentiviral Vector System, Viral Gene Therapy,... -
Tips for a 1st Time CRISPR User (by a 1st Time CRISPR User)
TypeBlog PostPublishedMarch 7, 2017, 3:30 p.m....Stable Cell Lines with Lentivirus Isolating a Monoclonal Cell Population by Limiting Dilution Lentivirus...high Cas9 MOI considering that A549 cells are relatively easy to transduce. I ended up with too many Cas9...be uniform editing across all the cells in the population, so some of the cells will still appear green...efficiently editing, don’t expect a 100% knockout population. BRAF gRNA delivery Overview: In practice you...on genomic DNA. The surveyor assay itself was relatively easy. Figure 4: Detection of edited BRAF...get an idea of the percentage of cells in the population that have an edit. I didn’t do this, but to do... -
Live and Let Dye: Self-Labeling Protein Tags
TypeBlog PostPublishedApril 16, 2024, 1:15 p.m....BG) (Keppler et al., 2003). This engineered hAGT later became known as SNAP-tag. There is also a fast-reacting...demonstrated in 2005 that proteins fused with E. coli dihydrofolate reductase (eDHFR) could be labeled with fluorescent...Although this tight interaction is not covalent, later work showed that introducing a Leu28Cys mutation...SNAP-OMP25 with Atto590-BG (green), imaged with stimulated emission depletion (STED) nanoscopy. Scale bar...different ligands to the same sample to label sub-populations of your protein of interest. For example, the...-molecule tracking of actin filament assembly-regulating proteins in vitro reveals complex events. a) ...Gaska, I., & Shekhar, S. (2023). Multicomponent regulation of actin barbed end assembly by twinfilin, formin... -
Hot Plasmids - September 2019 - CRISPR, Auxin Inducible Degradation, MerMAID Channelrhodopsins, and Genome Looping
TypeBlog PostPublishedSept. 3, 2019, 1:15 p.m....technology (Yesbolatova et al., 2019). Find the mAID tagging toolkit at Addgene Yesbolatova et al., Methods...ubiquitin ligase, the Kanemaki lab was able to translate AID to yeast, as well as mammalian and human cell...tagging of proteins with mAID and other tags (Yesbolatova et al., 2019) using CRISPR. This toolkit makes...thought to play a key role in the transcriptional regulation of many genes. A number of groups have designed...synthetic looping factors developed thus far for these latter experiments, such as transcription factors tethered...