...fluorescent proteins system for Caenorhabditis elegans By Jennifer Tsang Fluorescent proteins (FPs) traditionally...Tools for labeling excitatory inhibitory synaptic proteins Fluorescent protein system for C. elegans Fluorescent... Labeling excitatory and inhibitory synaptic proteins with FingRs By Jason Nasse Since the pioneering...neurotransmitters. Using genetically encoded fluorescent proteins coupled with Fibronectin intrabodies generated...FingR constructs target the synaptic scaffolding proteins PSD95 and gephyrin to label excitatory and inhibitory...neuron, the FingRs bind to the endogenous synaptic proteins allowing them to localize to the synapse and minimize...combining vectors expressing different fluorescent proteins targeting inhibitory or excitatory synapses or...

... process. When using zinc finger proteins or TALE proteins, proteins had to be re-engineered for each ...that bind to MS2 proteins. These MS2 proteins then recruit additional activation domains (HS1 and p65). ...transcriptional activators: some methods rely on fusion proteins while others re-engineer components of Cas systems...method is its construction: it relies on antibody chains, which are relatively large and are not expressed...epitope tag of SunTag with the P65-HSF activation domains used in SAM. This method has been shown to activate...Davidsohn N, Housden BE, Perrimon N, Weiss R, Aach J, Collins JJ, Church GM (2015) Highly efficient Cas9-mediated...Kowal EJK, Buchthal J, Housden BE, Perrimon N, Collins JJ, Church G (2016) Comparison of Cas9 activators...

...images of proteins. But they are still flat. To truly appreciate the 3D structure of these proteins, it is...Molecular Designs, we create physical models of proteins using 3D printing technology. The tagline for ...to create stunning, interactive images of their proteins. Since the 1980’s, journal articles have been ...structure? We have been creating physical models of proteins for researchers for the past fifteen years. On...expressed purpose of creating physical models of proteins and other molecular structures for use in professional...Under the guidance of a trained educator, these instances of focused attention become magical teaching moments...their research while creating physical models of proteins that are being investigated in their labs. Soon...

...elements anymore. The kit contains sequences for several different fluorescent proteins. Visualizing fluorescence...Nicola Patron is Head of Synthetic Biology at the Sainsbury Laboratory, where she often feels more like an... between plants and their pathogens. As Patron explains it, her work has always been focused on gene transfer...Cloning (MoClo) Kit. What does it include? Patron: Sainsbury has a mandate to share and educate and enable ...chlorophyll, so we looked at newer fluorescent proteins that get around that problem. MoClo is really ...straightforward really and we’ll be producing an instructional video soon to make it easier for people to use...for plants? Patron: Our lab and others at the Sainsbury are working on a lot of CRISPR tools for editing...

...forces its insect host to dive into water. There, the worm emerges to find a mate, leaving the insect dead ...infected grasshoppers expressed different proteins in their brains compared to uninfected grasshoppers, including...Plasmid pumpkins, team costumes, and spooky science stories. Yes, please! At Addgene, we love finding...addition to the Halloween contest, we’ve also carved pumpkins (yes, Blugene made an appearance there too) and...base of the tongue where it pierces the tongue and drains all the blood out. Once the blood is depleted, ... the tongue atrophies and falls off. The louse remains becoming the fish’s new “tongue” (Brusca and Gilligan...Spinochordodes tellinii‘s larvae makes its home inside grasshoppers and crickets. When it’s ready to transform...

...lentivirus, but instead is contained by a capsid which is comprised of the viral proteins VP1, VP2, and ...Maday et al., 2014). It is currently unclear which proteins facilitate transport of neuronal tracing compounds...peroxidase (HRP; Kristensson and Olsson, 1971), plant lectins (wheat germ agglutinin WGA, Schwab et al., 1978...iontophoresis (applying small electric currents via inserted electrodes) may be applied, but these methods ...neuroanatomy and neural connectivity, providing valuable insights for neuronal communication. Viral vectors and...methods such as electrophysiological recordings. The origins of these methods are naturally occurring viruses...engineered to have differences in their capsid proteins. For a brief overview see Addgene’s AAV guide....

...tweet URLs with scientific domains <10% of the time. Figure 3: Top 20 Domains Tweeted by Scientists. 6....scientific domains more often than other scientists   Figure 5: Portion of Tweets with Scientific Domains, Categorized.... The #1 (re)tweeted domain by scientists was Instagram Social media and news sites were also popular....6. The top scientific domains tweeted by biologists were: Nature, Sciencemag, Wiley, Cell and Scientific...Scientific American Figure 4: Top Scientific Domains Tweeted by Biologists. 7. Biologists, neuroscientists ...(orange box) post tweets containing scientific domains more often than other disciplines. The majority...tweets for most disciplines contain scientific domains between 0-10% of the time; however, ~20% of life...

...encoding the insert of interest as well as sequences necessary to encapsulate that insert into a viral... psi signal, the genes encoding the structural proteins could not be integrated into the new viral particles... one with the exception that the HIV accessory proteins that are not essential for the production of the...its activity. A CMV or an EF1α promoter has been inserted in this LTR so you don’t need Tat to transcribe...RCL) Efficient gene transfer Potential for insertional mutagenesis: Replication-incompetent lentiviruses...human tropism can still infect the researcher and insert into her/his genome - this is a biosafety risk ...dividing and non-dividing cells No immunogenic proteins generated Stable integration into host genome...

...editor can install all possible point mutations, delete short stretches of DNA and even insert new DNA at...advancements and the noteworthy work of fiction that it inspired. CAUTION: Spoilers alert! Pre-DNA Era  H.G. ...enzymes, meganucleases, were discovered. These proteins are characterized by large DNA recognition sites... successful in the box office, indicating the insatiable appetite of the moviegoers for dinosaurs wreaking... In this movie Ethan Hawke’s character revolts against class divide that is created by genetics.    Zinc...DNA-cleaving strategy similar to the ZFNs, but instead of recognizing nucleotide triplets, the components...target new genomic sites. The discovery of the Cas proteins in 2005 was a major breakthrough in the world ...

... is present even in common lab cloning strains. This intrinsic bacterial recombination pathway, referred...region (calculated using Oligo Calculator). Insertions - To insert short sequences of up to 100 bp, such as...referred to as recA-independent recombination, joins together pieces of linear DNA through short homologous...recombination reported in all laboratory E. coli strains tested so far. The use of this pathway as a cloning...the PCR mix to remove parental DNA (37 °C for 15 mins). This enzyme specifically cuts methylated DNA, ...used to perform any plasmid modification, from insertions, deletions, and point mutagenesis to complex ...template binding regions to amplify away from the insertion site. Encode half the new sequence at the 5’ end...

...Director of the Merkin Institute and Vice-Chair of the Faculty at the Broad Institute; Professor of Chemistry...Harvard University, and David R. Liu from the Broad Institute and Harvard University. Over the course of the...papers (data was provided by Addgene on 5/26/20 and contains requests from 5/22/14 to 5/19/20). Each paper ...data with a 7-day rolling mean gave us a crude insight into the magnitude of the effect COVID-19 had on... analytical expression for the data likelihood contains an intractable integral of the product of a Poisson... least impacted. These results highlight the instability and uncertainty in estimating parameters from...important additional caveat is that our dataset contains plasmid orders from just one lab, which limits...

... antibody against PARIS, a zinc finger protein involved in neurodegeneration in Parkinson’s disease. Knowing...rather than producing and purifying it in your lab. Instead of starting a new search or clicking through to...when we changed from searching for viral preps instead of plasmids? The available filtering options now...types of searches including depositing labs and institutions. For example, search for “Melina Fan” and you...Flame is more than an award for depositors and inspiration for our updated mascot, it’s also a convenient...Species: Specify the source organism for a gene insert. (e.g. S. cerevisiae) Plasmid Type: Choose an empty... empty backbone if you intend to clone your own insert, or use the other options to find plasmids matching...

...to modified proteins called histones. We can detect modified histones with “GPS for proteins,” ChIP-seq... coordinates: chromosome, start, and stop. For instance (chr3, 1, 10) indicates a stretch of DNA at the...techniques Technique What It Locates ChIP-seq Proteins (for our example later, histones) ChiRP-seq ...For details on DNA compaction and the structural proteins that make it possible, see the appendix DNA is... you run a ChIP-seq experiment (think “GPS for proteins”) for NANOG, an essential transcription factor...epigenomics: H3 – one of NANOG’s associated histone proteins K4 and K27 – locations of the amino acid lysine...enhancers, lncRNAs, repetitive elements, repressors, insulators, and more. We know that this matter exists, but...

...locus-specific ChIP). These methods consist of insertional chromatin immunoprecipitation (iChIP) and engineered...interactions. For this purpose, we developed the insertional chromatin immunoprecipitation (ChIP) (iChIP) ...purification, in 2009. However, iChIP requires insertion of recognition sequences of an exogenous DNA-binding... DNA-binding molecule technologies such as TAL proteins and CRISPR became available, I was very excited...easily isolated for identification of associated proteins, RNAs, and other genomic regions. I believe that...genome functions in the hope of developing drugs against intractable diseases. Currently, a number of labs...analyze signal-induced nuclear translocation of proteins in 2004. In addition, we have recently developed...

...lines. Identifying less toxic envelope proteins or envelope proteins that target specific tissues are areas... lentiviral expert) Didier Trono's lab webpage explains how lentiviral particles are produced by co-transfecting...that's gag, pol, and env), the isoforms of these proteins, as well as the viral long terminal repeats (LTRs...envelope plasmid and a packaging plasmid encoding HIV proteins gag, pol, rev, and tat. 3rd generation systems...In addition the deletion reduces the risk of insertional mutagenesis by limiting the interaction between...components are HIV-based? A7: No single plasmid contains all the components necessary to produce viral ...vectors contain gag, pol, and rev. Second-generation contains those three genes plus tat. The envelope plasmid...

...Place intended insert in between nick sites Use 40 nt homology arms for small insertions or tags (ssODN...By mutating one of two Cas9 nuclease domains, researchers created the CRISPR nickase. Nickases create... creates a non-target strand-cleaving nickase. Instead of cutting both strands bluntly with WT Cas9 and...deletions, while H840A is biased towards large insertions. These profiles are likely due to the distinct...can’t find a good gRNA that cuts close to your insertion site, you can’t obtain high HDR efficiency. Nickases...and Yan designed a PAM-out nickase experiment to insert an EcoRI site using a single-stranded oligonucleotide...oligonucleotide (ssODN) donor. Using a D10A nickase to insert an EcoRI site in various spots between the nick...

...segment of DNA which contains several restriction sites allowing for the easy insertion of DNA. In expression... Produce proteins that glow so that scientists can track their location or quantity inside a cell ...2014 and updated Apr. 14, 2020. Any newcomer who joins a molecular biology lab will undoubtedly be asked... a plasmid by recruiting replication machinery proteins Antibiotic Resistance Gene Allows for selection... the MCS is often downstream from a promoter. Insert Gene, promoter or other DNA fragment cloned into...commonly referred to as “vectors” or “constructs.” To insert a gene of interest into a vector, scientists may... are complete, the vector containing the newly inserted gene is transformed into bacterial cells and selectively...

...proper folding of the tag, which prevents insoluble fusion proteins from being purified by GST (maybe try ...between – now all you need are some of your favorite proteins purified to carry out your plan. With a little...purification a cinch. Types of affinity tags, the proteins they work best attached to, and the species they...will be expressed along with all other cellular proteins within the expression system (bacteria, yeast,...tags also have commercially available antibodies, resins, beads, and other purification aids readily available...for detection and/or purification of MBP-tagged proteins. The antibody is an alternative to amylose resin...Nickel, Cobalt, Copper, etc.), allowing His fusion proteins to be purified by single step metal affinity chromatography...

...vague and "what proteins regulate the Z pathway?" is an open-ended list of proteins. Instead, consider "how... at the National Institutes of Health, was an Associate Professor at Albert Einstein College of Medicine...Hughes Medical Institute. His interests include the quality control of secretory proteins in the Endoplasmic... Research Campus of the Howard Hughes Medical Institute. Are you currently on or planning to go on the..."how do proteins X and Y regulate the Z pathway?" A well-defined outcome makes it easier for the search...favor a different model. Finding evidence for or against a model is a standard scientific strategy. However...Endoplasmic Reticulum, optimization of fluorescent proteins,  and live cell imaging approaches. Erik is also...

...used for an array of detectable proteins including fluorescent proteins, antibiotic resistance, and luciferase...monitors APOBEC (a family of cytidine deaminase proteins) and Cas9-mediated editing in real time (St. Martin...inactive mCherry, the lab introduced a 43 base-pair insertion that disrupts fluorescence due to a frameshift...TCA-to-TUA) found within the 43 base-pair mCherry insertion. Editing of these motifs generates lesions that... editing capability of the seven human APOBEC3 proteins, finding that APOBEC3A and APOBEC3bctd are the...populations. To develop this method the lab was inspired by work that used an integrated GFP reporter that...DNA editor. Specifically the BFP mutant (BFPH66) contains a histidine at amino acid 66. This histidine is...