We narrowed to 854 results for: REN
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TypeBlog PostPublishedJuly 15, 2014, 4:03 p.m....than 300 different coactivator proteins individually contribute to cell regulation in a coherent manner.... one of the most successful targets for drugs currently available or being developed to treat a multitude...patterns of nuclear receptors across multiple different tissues in normal and disease states, their circadian...previously uncharacterized cellular roles. In the current phase of NURSA we are seeking to compliment these...our data mining tool, Transcriptomine. We are currently nearing the completion of an 18-month effort to...found in the Datasets and Reagents sections of the current NURSA website. NURSA and Addgene The great progress...
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Viral Vectors 101: Calcium Sensors
TypeBlog PostPublishedAug. 10, 2023, 1 p.m....remember: Keep Ca+2lm and Ca+2rry on! References and Resources References Nasu Y, Shen Y, Kramer L, Campbell...sensors are commonly used calcium reporters with different kinetic parameters. There are three GCaMP types...and Localization The calcium concentrations in different subcellular compartments can vary greatly. Additionally...measurements are more susceptible to artifacts and pH interference, and comparison between experiments can be more...difficult due to variance in sensor expression if you aren’t working with a stably integrated biosensor. Ratiometric... also suitable for in vivo studies. There are different versions optimized for neuron-specific, astrocyte-specific... -
How to Prepare for an Industry Interview
TypeBlog PostPublishedJuly 9, 2020, 1:15 p.m....job hunt, I quickly noticed one of the biggest differences between applying for an academic position versus... phone interview. I also transitioned between different lab groups without having a formal interview process...usually have a theme too, which is something not inherently captured by the STAR method. I like to think ... try and circumvent this, test out the video conferencing software you’ll be using in advance of the interview...interview, if possible. Otherwise, read the video conferencing instructions you received from HR carefully....completing each section of your talk. Some video conferencing software allows participants to “raise” their... wouldn’t have made it to the interview if you weren’t qualified for the position. So prepare for the ... -
Synthetic promoter AAVs for cell-type specific expression in retinal cells
TypeBlog PostPublishedMarch 12, 2020, 1:15 p.m....their current promoters, as well as more promoters that target cells types missed by their current library... Clinical Ophthalmology Basel recently took a different approach to enhance specificity in retinal cells... synthetic promoters, the Roska lab used four different design strategies: Strategy #1 (ProA): Create... this library contained 226 AAVs, each with a different synthetic promoter driving expression of the same...was from a single class of cells Of the four different promoter designs, promoters from strategy #4 had...ready-to-use AAV viral preps Browse all viral vectors References Juettner, J., Szabó, A., Gross-Scherf, B., Morikawa... -
A Look Back at One Year of Plasmid Sharing for COVID-19 Research
TypeBlog PostPublishedJuly 12, 2021, 1:15 p.m....never been so important. References and resources References Biering SB, Sarnik SA, Wang E, Zengel...for generating pseudovirus and those encoding different spike protein variants that can be incorporated...vaccinated individuals are at neutralizing the different SARS-CoV-2 spike variants. You can learn more ...head over to this blog post that highlights five different methods to use CRISPR to detect SARS-CoV-2. Other... including those from Andrea Pauli and Julius Brennecke. They developed a method that uses loop-mediated...Whiteman MB, McKay LGA, Lelis FJN, Habibi S, Cai Y, Rennick LJ, Duprex WP, McCarthy KR, Lavine CL, Zuo T, Lin... -
BeHeard Award 2018: Diseases of Glycosylation, Arginine Mutagenesis, & Neural Development
TypeBlog PostPublishedJune 26, 2018, 1:33 p.m....life-threatening consequences. They result from defects in different steps in the glycan biosynthetic pathway and have...characterized by a predisposition to severe and recurrent infections along with a mild or exacerbated inflammatory...infection and other pathological conditions. References: Monticelli, M., Ferro, T., Jaeken, J., dos Reis...multi-subunit macromolecular complexes, but a serendipitous discovery led them to investigate the use of... discovered that the in vitro functions of 11 different Arg-mutant enzymes could be rescued by supplementation...diseases caused by Arg-mutant proteins, but there is currently no evidence of Arg-mutant protein rescue in living...from rare diseases arising from Arg mutations. References: Baldwin, Enoch, et al. "Generation of ligand... -
Nanoblades: Tiny CRISPR Ninjas for Genome Editing Difficult Cells
TypeBlog PostPublishedSept. 26, 2019, 12:50 p.m.... 1-4 different gRNAs. Viral envelope proteins To alter the cell tropism of Nanoblades, different viral...incubating Nanoblade particles with polybrene, a cationic polymer. Polybrene has long been used to increase...electroporation, and viral transduction; but these methods aren’t stealthy enough to gain access to some cells and...embryos, and animals. Nanoblades can also carry different types of CRISPR cargo including RNPs for indel...versatile and accessible genome editing tool. References Mangeot, Philippe E., et al. "Genome editing ... -
Fluorescent Proteins 101: Fluorescent Protein Timers
TypeBlog PostPublishedMay 4, 2017, 2:30 p.m....but timer activity can be calibrated to these different conditions, enabling the use of a given timer ...While Tsuboi et al developed mK-GO somewhat serendipitously while attempting to enhance mKO in other ways... of Neurog3 (a gene controlling pancreatic differentiation) in mouse embryos (Miyatsuka et al.) Distinguish... Let us know in the comments section below! References 1. Terskikh, Alexey, et al. ""Fluorescent timer...11090358. 2. Tsuboi, Takashi, et al. "Age-dependent preferential dense-core vesicle exocytosis in neuroendocrine...and Michael S. German. "Chronology of islet differentiation revealed by temporal cell labeling." Diabetes...programmable synthetic lineage-control network that differentiates human IPSCs into glucose-sensitive insulin-... -
Truncated gRNAs for Regulating Gene Expression
TypeBlog PostPublishedJan. 10, 2017, 3:37 p.m....have proposed the concurrent use of “orthogonal” Cas9 proteins derived from different bacterial species...variants—nuclease-competent or nuclease-null—in order to effect different perturbations. Consider, for instance, the challenge... both cutting and regulation of expression at different genetic loci within a single cell. One might imagine...expression (6). Altering the length of gRNAs targeting different sites of interest thus represents a straightforward...introduced together with truncated gRNAs targeting a different set of loci to induce simultaneous cutting and...DNA sequence or regulating gene expression. References 1. Esvelt, Kevin M., et al. "Orthogonal Cas9 ... -
Tips for a 1st time AAV user (by a Rookie AAV user)
TypeBlog PostPublishedOct. 23, 2018, 11:49 a.m....into the margins of my protocol. While these tips weren’t critical to the experiment, they definitely made... AAV packaging plasmids. The cells need three different plasmids to produce AAV: 1) the RepCap plasmid...gradient ultracentrifugation uses a gradient of different concentrations of iodixanol to separate out contaminants...titer generated by qPCR is useful, I test a few different concentrations of virus, or multiplicity of infections...variability. Check out this post to learn more about different ways to titer AAV. Do you have any AAV production...teaching me about AAV production and purification! References 1. Aurnhammer, C., Haase, M., Muether, N., Hausl...adeno-associated viral vectors for in vitro and in vivo use. Current protocols in molecular biology, Chapter 16, Unit... -
Transferable Skills Guide: Managing a Team
TypeBlog PostPublishedJan. 30, 2018, 2:42 p.m....test: my undergrads were different people! More specifically, they had different learning styles. One was... can be very different from what motivates you!), know their unique skills and strengths (you will likely...likely have reports with different levels of experience), find out what their learning styles are and capitalize... to you to decide which one you want to be! References Becoming the boss. Harvard Business Review, ... Karen Guerin... -
Transgenic Organisms, Cas9 Gene Drives, and Appropriate Safeguards
TypeBlog PostPublishedMay 22, 2015, 4:58 p.m....safely and transparently. For more information, visit www.sculptingevolution.org. References: Burt, ...didn't work quite as well as anticipated4, greater awareness will certainly improve safety. What safeguards...vectors of diseases such as malaria and dengue are currently being performed in regions that don't have resident...Cas9 genome editing. Finally, gene drives are an inherently collective technology: because they necessarily...dengue for many years. Since more than a thousand children died of malaria today, let's not risk a potential...information about gene drives can be found in the referenced publications. Thank you to our guest blogger... -
22 Hot Plasmid Technologies from 2014
TypeBlog PostPublishedJan. 6, 2015, 5:21 p.m.... mutagenesis or gene targeting Victor de Lorenzo's lab has engineered a modular mini-Tn5 vector that... microscope 8 hours post-pulse. The distinct differences of red-to-green fluorescence in individual mitochondria...mitochondria in the networks marks the differences in import of newly synthesized (green) protein and ...five previously unknown channelrhodopsins from different species: Stigleoclonium helveticum (ShChR), Chlamydomonas...cassette. They validated this methodology in two differents types of bacteria (E. coli and Salmonella typhimurium... of all four final RVD modules in each of two different TALE scaffolds are provided, as the optimal scaffold...region of one neuron and postsynaptic region of a different neuron, come into proximity in the synaptic cleft... -
CRISPR 101: Multiplex Expression of gRNAs
TypeBlog PostPublishedSept. 10, 2020, 11:45 a.m....delivered on a different plasmid and allows simultaneous multiplex editing of six different genes. This ...flanking the promoter-gRNA construct, but with different sequences adjacent to the sites. When digested...assembly principles, but they’re optimized for different purposes. Gersbach Lab multiplexing plasmids...kanamycin-resistant plasmids, each containing a different gRNA target sequence downstream of the 7SK, human...users to express up to 7 gRNAs. The kit contains different destination vectors depending on the total number...transcript. Rather than being transcribed from different promoters, the gRNAs are transcribed together ...likely be used in a wide range of bacteria. References 1. Maddalo, Danilo, et al. “In vivo engineering... -
Antibodies 101: Producing Recombinant Antibodies
TypeBlog PostPublishedAug. 13, 2024, 1:15 p.m....space to be oxidized and form disulfide bonds (Frenzel et. al., 2013). If you’re using a mammalian production...: The steps of rAb production. Created with biorender.com Another method of harvesting uses magnetic...useful to compare recombinant antibodies to their parental hybridomas. Aggregation and storage Aggregation...our Antibodies 101 blog series. Resources and references More resources on the Addgene blog Introduction...Purification Protocol Addgene's Antibody Guide References Trimmer JS. Recombinant Antibodies in Basic Neuroscience...: 10.1007/978-1-4939-8730-6_1. PMID: 30242676. Frenzel A, Hust M, Schirrmann T. Expression of recombinant...particle formation, characterization & rheology, Current Opinion in Colloid & Interface Science, Volume ... -
Negative Can Be Positive: Open AAV Data with Addgene
TypeBlog PostPublishedSept. 6, 2022, 1:15 p.m....Testing of three different AAV serotypes (AAV1, 8, 9) injected into the A2 noradrenergic area of the caudal...helping everybody, helps everybody! References and resources References Dayton, R. D., Grames, M. S., & ...scientists under-informed with respect to how different serotypes, promoters, and injection methods are...literature, scientists can use the Data Hub to compare different methods and vectors, and use this data to guide...particle aggregation and methods to prevent its occurrence during vector purification and formulation. Molecular... -
AAVs CREATed for Gene Delivery to the CNS and PNS
TypeBlog PostPublishedSept. 28, 2017, 2:01 p.m....be identified by PCR later. The PNS presents a different problem: it’s spread throughout the body and is..., AAV.PHP.eB was used at a lower dose than it’s parent AAV.PHP.B: 1x1011 vs 1x1012 viral genomes (vg) ...comparison of AAV.PHP.eB and AAV.PHP.S to their parental vectors. To quantify this comparison, capsids ...efficiency of AAV-PHP.eB and AAV-PHP.S versus their parental AAV vectors. What can these AAV’s be used for... Cortex Striatum Cerebellum AAV.PHP.B (parental) 49 +/- 7% 36 +/- 8% 37 +/- 10% AAV.PHP.eB 69...Transduced Dorsal Root Ganglion (DRG) AAV.9 (parental) 46 +/- 0.7% AAV.PHP.S 82 +/- 2% Table 2:... and supplemental figure 8 in Chan et al). References 1. Chan, KY, Jang MJ, Yoo BB, et al. Engineered... -
AAVs in Retinal Gene Therapy
TypeBlog PostPublishedJuly 31, 2018, 12:55 p.m....It was by serendipity that I got into the field of gene therapy, more specifically AAV-based retinal ...treatment, famously visited Congress to help increase awareness about the potential of gene therapy!). It would...is available. While only natural serotypes are currently used in clinical trials, a variety of engineered... AAV safety profile is excellent and further strengthens its position as the vector of choice for in vivo...and the development of personalized medicine. References and further reading 1. FDA approves gene therapy...the Light after 25 Years of Retinal Gene Therapy. Trends Mol Med. 2018, S1471-4914(18)30123-0. DOI: 10.1016... Karen Guerin... -
Deep Dive: Fixing and Permeabilizing for Immunofluorescence
TypeBlog PostPublishedAug. 30, 2022, 1:15 p.m.... as fixing and permeabilizing. References and resources References Brown-Harding H (2020) Fixation artifacts...previously hidden within the protein. Created with BioRender. Permeabilization What is the point? Antibodies...100, Tween-20, or saponin. Each detergent has different chemical properties that will impact the level...concentration. Figure 2: Examples of different permeabilization strategies and when to use them...permeabilizing organellar membranes. Created with BioRender. How do I choose?! Ok, so now you know some...https://doi.org/10.1177/33.8.3894499 Im K, Mareninov S, Diaz MFP, Yong WH (2018) An Introduction to... -
Antibodies 101: The Four ELISAs and When to Use Them
TypeBlog PostPublishedJuly 25, 2024, 1:15 p.m....scientists have since adapted it for a host of different proteins and small molecules from a variety of...needs. Herein, we’ll give you a rundown of the different types of ELISA, advantages and disadvantages of...in the sample. This figure was created with BioRender.com Direct ELISAs A direct ELISA is the quickest...this method, the target antigen competes with a reference antigen for binding to a primary antibody. While...typically users first coat the multiwell plate with a reference antigen and then they preincubate the antigen-...allow the unbound primary antibody to bind to the reference antigen. The more antigen in the sample, the more... means less antibody available to bind to the reference antigen, producing a weaker signal. The major...