We narrowed to 605 results for: RAN-1

...updated version of BRET called BRET2. Figure 1: TRUPATH measures heterotrimeric G protein dissociation...-visual G protein signal transducers (collectively known as the transducerome) that can be activated downstream...Resonance Energy Transfer (BRET). BRET is similar to Förster Resonance Energy Transfer (FRET), in which...non-visual G protein transducers. To use the TRUPATH system, cells are transfected with plasmids expressing...including smell, taste, sight, inflammation, and neurotransmission. But with approximately 950 different human...involved in sight (Olsen RHJ et al., 2020). A transducer complex consists of a trimer of three subunits...such a tool is being able to identify specific transducer complexes that are activated in response to a...

...enables the high-resolution tracking of more than 1 million cancer cells under drug treatment.” Stegmeier...members to the Board of Directors: Khandan Baradaran, Frank Stegmeier, and Luk Vandenberghe. We’re excited...members and get to know them! Khandan Baradaran, PhD Khandan Baradaran’s expertise in viral safety, gene therapy... with health authority regulations,” Baradaran says.  Baradaran’s path to science began in high school... getting these reagents is profound.” Frank Stegmeier, PhD Frank Stegmeier is the Chief Scientific Officer...in viral genetics. In regards to open science, Baradaran thinks the best way to solve problems is to work...think will get the research funded,” she says. Baradaran met Addgene Executive Director Joanne Kamens several...

...brain where DA quantitation is desired (Figure 1). The probe perfuses liquid, which causes the passive diffusion...dopamine (DA), a well-known reward chemical and neurotransmitter. But aside from DA’s famous role as the “happy...standard method of measuring DA and other neurotransmitters in the brain has been in vivo microdialysis...performed with a probe surrounded by a semipermeable membrane, which is inserted into the region of the brain... diffusion of DA across the semipermeable membrane from the extracellular fluid into the perfused liquid... to a threatening looming stimulus in vivo in transgenic zebrafish Report DA release in live mice over...developments in the detection and measurement of neurotransmitters in various species and while exhibiting complex...

... to just that particular assay.      Fig. 1: A native protein (left) is denatured, allowing the...bound to anything? Is it a transmembrane protein, or is it know to translocate or fold in certain conditions...as part of performing a specific function. A transmembrane protein that is being used as a marker for flow...cell, instead of when it is attached to a cell membrane.  Picking the Right Antibody  It can get even ...looking for a protein of interest while it’s in a transition state or specific configuration. So how do you...

...vectors (e.g. pBV-Luc) and gene silencing (e.g. pLKO.1 - TRC cloning vector). Check out our complete guide...). If you can get away with generating transiently transfected cells, you will not need a mammalian selection...mammalian selection cassette to screen for positive transfected cells. The addition of an in-frame fluorescent...create a new strain with a disrupted gene by transforming wild-type yeast with an integrating vector expressing...majority of fly vectors are based on p-element, random or targeted, integration into the fly genome and...applications used to study gene function. From making transgenic animals, gene targeting and genome modifications...

...ScFvs, based on community demand. Figure 1: Antibody against PSD-95 from the Trimmer Lab. Information... our values of transparency, reproducibility, and community-building. The transparency begins with sharing...Addgene’s antibody resource is funded by a generous grant from the NIH’s Brain Research Through Advancing ...share plasmids with each other. Together, we can transform the antibody sharing landscape as well.     References... Improving Reproducibility through Rigor and Transparency of Experimental Methods. Neuron 90(3):434-6.... project is supported by a NIH BRAIN Initiative grant (U24NS119916 ). The content of this article is solely...

...materials to the research community. References 1. Coroadinha, A. S., et al. "Effect of osmotic pressure... AAV2 interacts with the packaging cell's cell membrane When packaging adeno-associated viral vectors ...to be due to the viral particle’s affinity for heparan sulfate proteoglycans on the surface of the packaging...supernatant, the viral particles associate with the cell membrane and are taken into the cell via endocytosis. In...looking to improve their AAV yields. Given the wide range of downstream applications scientists rely on our...preparation. Through sugar detection assays, in vitro transduction, and silver stain approaches we show that there...test the endocytosis of a fluorescent dye and transduction of GFP-expressing AAV particles we did not observe...

...the life scientist or bioengineer.   Figure 1: Map of a plasmid with its elements described below... origin of replication (which controls the host range and copy number of the plasmid) and typically include...for further study. Promoter Region Drives transcription of the target gene. Vital component for expression...cell types. Primer Binding Site A short single-stranded DNA sequence used as an initiation point for PCR...vector containing the newly inserted gene is transformed into bacterial cells and selectively grown on...biologists to broadly utilize plasmids across a wide range of applications. Some common plasmid types include...

...plasmids. The Zhang lab, on the other hand, uses a 1 plasmid, single infection system. Genome wide mouse...target cells that have previously been stably transfected for Cas9 expression. The Yusa lab has used their...

...literature? Very often the data of interest are in 1 or 2 figures from one paper and then another figure...preprints, data, software, micropublications) in grant proposals. ReFigures are one such research product...

... email in a busy inbox. Updated policy As of July 1, 2025, we are updating our Hold for Publication policy...materials are both excellent ways to provide transparency, support reproducibility, and accelerate science...

...could be kept frozen after collection. Figure 1: Paul included a voltmeter on top of the battery box...ahead of time included contacting YNP staff to arrange permits to visit places not accessible to the public...night was fun but also very tiring. It feels a bit strange to be using a micropipette and gloves at a campsite...similar to a type II-C Cas9 from Parvibaculum lavamentivorans, whose optimal growth temperature is 30°C. ...rRNA in bacterial RNA-Seq. This enriches for transcripts of interest without an increase in the number...

...sequencing the unique DNA barcodes. Figure 1: Schematic of barcoding with a pooled library. Created...Variations of this library allow tracking of transcription or chromatin accessibility at the single-cell... however, nuclease-dead Cas9 is fused to a transcriptional activator or repressor to create a reversible...Zhang lab developed this library of all human transcription factors, including splice isoforms, to allow...ExpRession Element. This library consists of broad host range regulatory sequences ready to be assembled with ...

...into three main effector functions.  Figure 1: Fc effector functions in an immune response to a ... the pores being opened in the pathogen’s cell membrane (Dunkelberger & Song, 2010). As cells cannot survive... survive when holes are punched in their membranes, the pore'd pathogens quickly lyse and die.  Antibody...where the eosinophils release cytotoxins via degranulation.  Antibody-dependent cellular phagocytosis (...

...do researchers track fluorescently labeled cells? 1. Epifluorescence microscopy This technique uses a ...of the immune system and its dynamics. References 1. Peters, Nathan C., et al. "In vivo imaging reveals...for a growing organism to make the complicated transition from a clump of cells to a fully differentiated...cooperate to cause a higher-energy electronic transition in a fluorescent molecule (see figure to the ...past 15 years efforts have been made to create transgenic mice expressing fluorescent protein reporter ...microscopy enabled scientists to see how chaotic and random the scanning was and how fast T cells could jump...Blue-green fluorescent proteins like EGFP Yellow-orange fluorescent proteins like TagRFP, tdTomato, DsRed...

...targeting MNase to antibody bound proteins (Figure 1). Importantly, since Protein A has weaker affinity...sonification. Though it helps capture weaker or more transient DNA-protein interactions, the reliance on fixation...McKay lab we study how differential access to transcriptional regulatory elements is controlled during development...

...in online patient/caregiver communities  Table 1: Examples of defining your audience for science communication...  For example, while “PCR test” has recently transitioned from jargon to non-jargon in the context of ...number of characteristics. Common ones include age range, familiarity with the subject, shared interests,...

...year and happy reading! Top 10 blog posts of 2015 1. CRISPR 101: Homology Directed Repair Addgenie Chari...used to generate inversions, deletions, and translocations. It also provides researchers with many different...Where Do I Find One? If you need help making a transition into a new job or maybe to an entirely new career...

...Rechsteiner M. Cell. 1982 May;29(1):33-41. PubMed. Protein transduction: unrestricted delivery into all...protein Tat. Although CPP fusions can translocate a cell membrane, the presence of the CPP may alter protein... (78-79% after two rounds of transduction.) Compared to CCP transduction, iTOP is at least four times ..., and cell death post-transduction is low. Neuronal stem cells transduced with iTOP resulting in...lab suggests that primary cells may be better transduced using only protein. Read on for a description...description of the lab’s iTOP protein-only transduction method and its potential applications to CRISPR/Cas9 genome...genome editing.   Protein-only transduction? Niels Geijsen’s group in Utrecht wanted to pursue a nonviral...

...double-stranding step and reduces both the time and cost for AAV genome sequencing.  Figure 1: The...previously require a preliminary double stranding step. The double stranding step was assumed to be necessary...genome is single-stranded and both TA-based ligation and tagmentation require a double-stranded template.  ...extraction, the [+] and [-] strands might naturally associate forming a double-stranded species that could be...extract is a heterogeneous mix of single-stranded and double-stranded species. While not all of the molecules...extract associate with complementary strands, enough double-stranded species exist in the mix to serve as...viral genome We hypothesized that this double stranding step might not actually be necessary. Since the...

... double-stranded breaks and can deliver large sequences and even entire genes. Figure 1: Mechanism...affiliated TniQ protein recruits transposition cofactors. The transposon proteins integrate a payload sequence... like we finish writing about one dual CRISPR-transposon system before another exciting new advance emerges...combined with the large sequence capacity of transposons open whole new worlds to explore. The newest ...called evoCAST — an evolved CRISPR-associated transposase (CAST). Back up a second. What are CASTs? CASTs...bacteria. In the course of evolution, Tn7-like transposons captured and repurposed nuclease-deficient CRISPR-Cas...Cascade shown) combines with a donor template and transposon proteins (Tns) to integrate large sequences into...

... Complex Synthetic Transcriptional Programs with CRISPR RNA Scaffolds. Cell, 160(1), 339-350 (2015). PubMed...recent paper, "Engineering Complex Synthetic Transcriptional Programs with CRISPR RNA Scaffolds." Adam has...visualize the concepts discussed in the paper. The transcriptional control of multiple loci is deftly coordinated...cerevisiae with few techniques that are easily transferrable across host species or metabolic pathways. A...CRISPR-based RNA scaffolds to mimic natural transcriptional programs on multiple genes. Modifying the ...synthetic RNA scaffolds that can recruit various transcriptional regulators (Zalatan et al. 2015).  These scaffolds...nucleotide sequence then subsequently recruit a transcriptional regulator to modulate expression at that locus...

...heat-fixed smear with the crystal violet stain for 1 minute. Rinse the slide briefly with tap water. Flood...diderms, have a thinner peptidoglycan membrane and an outer lipid membrane. Common gram negative bacteria include...within their membrane, while the stain is washed from the gram negative bacteria’s thinner membrane. Procedure...destroys the outer lipid membrane, and the thin inner peptidoglycan membrane is unable to retain the stain...expanded on Gram’s discovery, adding a second stain, safranin, as a counterstain to help visualize the bleached...or monoderms, have a single thick peptidoglycan membrane. Common gram positive bacteria include staphylococci...and immediately rinse. Flood the smear with the safranin counterstain for 30 - 60 seconds. Rinse the slide...

... Figure 1: Identification and enrichment use of XMAS-TREE. a) Cells are transfected with three different...ready-to-use viral preps   Improved cell-free transcription/translation system to examine biosynthetic pathways... Gabrielle Clouse Recently, cell-free transcription/translation systems (also referred to as TX-TL systems...modifications to edit DNA without requiring a double-stranded break. The Xiao Wang Lab has recently deposited.... These plasmids use “Cas9-mediated adenosine transient reporter for editing enrichment”, or XMAS-TREE...identification of essential MYC binding sites in a wide range of human cells.  A new pooled library from Junjie...

...Tetbow (Tetracycline trans-activator Brainbow) (Sakaguchi et al., 2018) (Figure 1). Once introduced, fluorescent...fluorescent protein (XFP) genes in a transgene. When multiple copies of the transgene cassette are introduced, stochasticity...fluorescent proteins is only possible at low copy number ranges, resulting in low fluorescent protein level. The...gene delivery is more common than conventional transgenic strategies. In such vector-based gene delivery...in the original method, we used the Tetracycline trans-activator system to boost the expression levels,...expression level of XFPs, most likely by suppressing transcription. It is critical to express a minimal amount ...Bio-Protocol 8 (2018): e3046. Livet, Jean, et al. "Transgenic strategies for combinatorial expression of fluorescent...

...construction of redCALWY-1, a red-shifted FRET sensor for zinc using variants of mOrange2 and mCherry as donor...field is a series of fluorescence resonance energy transfer (FRET)-based calcium biosensors. Perhaps most ...developed and published in 2010 for visualizing the transcription factor cAMP-responsive element-binding protein...really important and when? That is not known.” Transition Metals Maarten Merkx of Technische Universiteit...particularly for the intracellular imaging of transition metal ions, such as zinc, copper, and iron. “...the sensors involves plenty of trial and error. Transition metals also present challenges in that they are... way to make a FRET sensor with a large dynamic range,” he says. Merkx recently applied this “trick” to...

...preferentially). You can also rank the plasmids by abundance, scaled to 1, to create a “cumulative wealth... seems straightforward: E. coli bacteria are transformed with the library DNA, they grow and replicate... Each Addgene library has its own depositor transformation and amplification protocol, but the overall...Maintain representation of the library during transformation First, use enough DNA and competent bacteria...and we use electroporation to ensure a good transformation efficiency. For a typical library, Addgene ...growing the cells on solid plates, where each transformant forms its own colony, uninfluenced by the growth...concern, but Addgene still recommends plating the transformation mixture on solid media. The colonies are then...

.... You can email her at [email protected].   References 1. Huang, Ally, et al. "BioBits™ Explorer: A modular...that green fluorescent protein through transcription and translation. Unfortunately, having live cells in... like gel electrophoresis to separate DNA or transforming cells with green fluorescent protein, these ... into a host cell (through a process called transformation), essentially giving the cell all the instructions...enzymes that can convert certain chemicals into fragrances that they can smell, or enzymes that can create...perhaps consider using some elements of BioBits to transition your undergraduate students from the classroom...

...occur, but good performers move around easily  Table 1: Taking it a Step Further - Big Company v. Small Company...encompasses millions of choices including pharma, tech transfer, management consulting, science communication,...into industry.” In his article On Leaving Academe, Terran Lane is asked by his colleagues and friends why...tell you to pick up the pace Constant worry about grants and renewals What is different about non-academia...outsource a task someone else who can do better, no grant writing Job changes can happen regularly (every ...where you want to go will help you focus on the transferable skills that you will need to get there. Additional...

... contain the full length plasmid.   Figure 1: The full lentiviral plasmid is expected to be >12...recombination, one resulting plasmid carries the transgene of interest (aka the viral genome) and the other...resistance gene and origin will replicate, and the transgene will be lost. Over time, bacteria containing the... grow! Our 5 Tips for Troubleshooting Viral Transductions will help with the next steps.   References...recombination of plasmid DNA without affecting mRNA translation efficiency or half-life. RNA 25:507–518. https...common lab E. coli strains Find tips for viral transduction  ...

...motion videos is very time consuming; producing a 1-2 minute video takes an entire week of our free time...have the same issues as one doing experiments in France. We can learn with each other through globalized...

...class.   Four ways to improve your next manuscript 1. Be consistent with your figures The figures of a ...worthless. But simply writing down your data doesn’t guarantee a publication. There is an art to composing manuscripts...

...wide range of input DNA concentrations. This process is explained more fully in the animated Fig. 1 below...plasmids on a plate) will typically have a wide range of input quantities. This variation is problematic...produced from large numbers of samples with a wide range of input concentrations (via proprietary chemistry...differences in input concentration across a 100-fold range (1ng – 100ng) are normalized to two-fold variation...

...heterodimerizes with CIBN upon blue light exposure (see Fig 1*). When the group targeted the enhancer region of ...ubiquitin ligase, the Kanemaki lab was able to translate AID to yeast, as well as mammalian and human cell...PubMed PMID: 31026591. Expanding the targeting range of CRISPRCas9 by engineering SpCas9 Article contributed...are also thought to play a key role in the transcriptional regulation of many genes. A number of groups...thus far for these latter experiments, such as transcription factors tethered to dCas9, require long time...

...then scroll on for some wintery delights!    Table 1: New ready-to-use viral vectors in the Addgene repository...Request service, which offers over 3,000 eligible transfer plasmids and five different serotypes. Not sure...

... and can likely be used in a wide range of bacteria. References 1. Maddalo, Danilo, et al. “In vivo...post about multiplexing with Cpf1.   Figure 1: Multiplexing allows researchers to express multiple...humanized wt Cas9, dCas9 (transcriptional repressor), and dCas9-VP64 (transcriptional activator)-containing...Multiplexing from a single transcript You can also multiplex gRNAs via a polycistronic transcript. Rather than being...being transcribed from different promoters, the gRNAs are transcribed together and are flanked by specific...simultaneously. Vectors for both transient expression and Agrobacterium-mediated transformation are available. For ...system for processing a continuous multi-gRNA transcript, each gRNA must be expressed from its own promoter...

...produce as many as 700 copies per cell. Figure 1: A plasmid map showing the standard features of a ...plasmids from the same group should not be co-transformed, so if you need two plasmids for an experiment... same incompatibility group should not be co-transformed. +Actual copy number varies. See below for ...stocks, agar stabs, or liquid cultures. Freshly transformed cells also give a higher plasmid yield that colonies...

...extension. Now, they split EB1 into two halves (Fig. 1): one containing the microtubule binding domain and...manipulations on cells, and has potential use for a wide range of applications in cellular biology. Van Haren ...fluorescent proteins (FP) with improved tissue penetrance and live cell imaging capabilities. Upon developing...

...Article contributed by Susanna Bachle Figure 1: Initial steps in the CRISPR-pooled screening using...Sabrina Büttner’s lab designed a calcineurin transcriptional reporter consisting of a quickly-degraded destabilized...contributed by Jennifer Tsang Figure 4: Double stranded DNA bound by tTALE-FP with the fluorescent protein...

...net negative charge with 4 negatively charged and 1 positively charged amino acids. Myc tags are frequently...some proteins. Protein folding issues, steric hindrances, and destabilization can be brought about by ...DOI: 10.1038/521274a Arribere, J. A., et al., Translation readthrough mitigation. Nature. (2016) 534, 719...

...defunct (but fun) plug-in called Maps.  Figure 1: See a starfish? This is actually a map of Joanne ...enough, but I’ll never forget the strawberries. Grand gestures are memorable! Attend events for one organization... want to do: Don’t connect via LinkedIn with strangers. It’s a waste of time. People that don’t know ...

...impartial. The best science happens when scientists have 1) the courage to make mistakes and 2) the integrity...of what I saw are that we, as a society, need to: 1) use more citations for out data so our sources are...I’ve worked tirelessly to get the publications, grant funding, and teaching experience that it takes to...

... lactose analogue, IPTG (isopropyl β-D-1-thiogalactopyranoside). (To be completely accurate, IPTG binds...x-gal (5-bromo-4-chloro-3-indolyl-β-D-galacto-pyranoside) into galactose and an insoluble blue pigment...α-peptide) to a lacZΔM15-mutant bacterial cell in trans complements the mutation allowing for a functional...site (MCS) into the α-peptide (represented as an orange wedge in the figure on the left) and inserted it...produces pigment from x-gal (included in the transformation plate medium), turning the bacterial colony...IPTG is also included in the medium to ensure transcription of the lac operon.   Tips for blue-white screening...screening Use a good control: Transform the backbone plasmid without insert. All colonies on this plate...

...red light rapidly deactivates the system. Version 1 of this far-red/red light-responsive system is spread...responses are varied; one outcome is directed transcription from a given promoter. To harness the power ...the RR (OmpR) via phosphorylation, promoting transcription (see figure below). Subsequent exposure to red...response regulator via phosphorylation, promoting transcription of a GFP reporter. Subsequent exposure to red...activity persisted in the inactive state. The dynamic range, defined as the difference between the lowest and... reduced system leakiness and increased dynamic range to around 100-fold. These improvements will allow...Applications of these systems The large dynamic range of these systems ensures that they can be used with...

...individual protein has all the components necessary to: 1. Bind to a guide RNA The guide RNA, which will be ...loci. 3. Cleave target DNA resulting in a double-strand break (DSB) Cas9 and its variants have two endonuclease...positions the nuclease domains to cleave opposite strands of the target DNA. Thus, the end result of Cas9...arrays as found in the bacterial genomes are transcribed into pre-crRNAs containing both the spacer region...RNase III, the tracrRNA and Cas9, bind to these transcripts and C) cleave them leaving mature crRNAs bound...target DNA which is E) cleaved leaving a F) double-strand break. A “gRNA” is a researcher-designed hybrid...

...through the use of nanoparticles. [6].    References 1. Zhang B. 2021. “CRISPR/Cas gene therapy.” Journal...editing system to correct the cystic fibrosis transmembrane conductor receptor (CFTR) by homologous recombination...Cas9 has been used to generate mutations in a wide range of organisms, but its potential to efficiently correct...Cas-based Gene Therapy in Human Genetic Diseases.” Theranostics. https://doi.org/10.7150/thno.43360 7. Yuxuan...

... will be a positive asset throughout your career? 1. Make scicomm practice a priority The good news for...next interview or whether your receive the next grant you apply for. After all, the only value your work...high level to their team.  When you're writing a grant, this means convincing a committee that your research...collaborating with colleagues in other fields or writing grant proposals to be reviewed by interdisciplinary panels...communities gather at ComSciCon's local and specialized franchise workshops throughout the year.  Find workshops...

...into account, the digest pattern depicted in Lane 1 would be expected for a XbaI/NotI digest. The program...DNA. The other component of these systems, methyltransferases, methylate DNA at particular sequences to...restriction modification systems containing methyltransferases that add methyl groups to specific DNA sequences...overlaps with the XbaI cut site (depicted as an orange line) because the restriction site is followed ...plasmids with no methylation (there are no methyltransferases in the PCR reaction). The template plasmid...

...challenges involved in plant synthetic biology. Challenge #1: Public perception of synthetic biology One of the...how they started their company here. The first transgenic plant was engineered over 30 years ago, but plant... still in its infancy. A long timeline from transformation to testing and a lack of well-characterized...s made even more difficult by the fact that transformation methods have also been patented. In order to.... We will also be working with open source, transformation technologies like those pioneered by Ewen Mullins...

... After all I was only going to be interviewed for 1 hour so having a broad enough insight was going to...promotions or further education may allow you to transition into various realms of business.  Thank you ...