...localization in cells, or even single-particle tracking of the bound complex. There are so many FPs available...fragments are named according to the number of strands in the canonical beta-barrel structure. Most frequently...frequently, the proteins are split between strand 10 and 11 to yield large (1-10) and small (11) fragments...in N-terminal fragment Constant rate of p53 tetramerization in response to DNA damage controls the p53 ...TagBiFC technique allows long-term single-molecule tracking of protein-protein interactions in living cells... can even be bright enough for single-molecule tracking in live cells.  Controls  When designing experiments...

...blot Run your gel and membrane transfer as normal. Immediately after transfer, dry the membrane to maximize...from having to run a whole new gel and membrane transfer. Let’s get started! Before you begin In the best...antibody binding to antigens. Ingredient Concentration or % Glycine or Glycine-HCl 25–200 mM ...more of your target protein. Ingredient Concentration or % Purpose Tris HCl 62.5 mM Buffer...whether that involves adding fresh chemiluminescent substrate or imaging your fluorescent probe. Then, if you...

...approved drugs so that your results are viable for translation to human patients. Picking receptors The receptor...nonnative ligands. However, they retain their intracellular interactions and still set off the same pathways... target effects. These synthetic agonists can penetrate the brain and some, such as varenicline, are already...Varenicline has been further modified to create ultrapotent PSEMS (uPSEMs) that have drastically increased... A, Basu J, Michaelides M, Sternson S (2019) Ultrapotent chemogenetics for research and potential clinical... specifically designed Gi-coupled receptor in transgenic mice. Nature Biotechnology 17:165–169 . https...

...China’s Ribobio, the illustrious speaker list attracted a host of sponsors based all over the world. With... market where one can shop for components of traditional Chinese medicine. There were hundreds of stalls...'ve overlooked regulatory fragments of these transcripts. I remember when small RNA was just something...’t give tRNAs much thought. High throughput transcriptome sequencing methods including work done by the...knew…I had barely heard of these). These single stranded, covalently closed RNAs seem to be conserved in...Using the Tet-on system, his lab has created a tetracycline cas9 inducible knock-out system. His lab is ...

...interactions (e.g., scores >80) are transformed to 100. This transformation (illustrated in the graph below) is performed...design tool Crispor by Max Haeussler. With a few extra lines of code, we enabled it to return guides and...the DNA target. CFD scores are then modified (“transformed”) such that weak interactions (e.g., scores <...re-create a match for the active guide. With these constraints in mind, we implemented the following algorithm...

...blastn), Standard Protein BLAST (blastp), and Translated BLAST (blastx). NCBI has a terrific getting-started...sequences (blastn)” program for blastn so that you can extract practically every single reliable base pair from...longer alignments to compare with your sequencing trace file. Unlike megablast, the regular blastn program...gaps in the nucleotide sequence, you can run a Translated BLAST (blastx) search to check for expected ORFs..., due to the additional searches involved in translating the nucleotide sequence into all six possible...choose for your sequencing result and can easily translate it, we recommend using blastp over blastx. The...

...or Förster] resonance energy transfer). FRET occurs when energy transfer from one FP (e.g. CFP) excites...the FP (fused to the protein of interest), thus tracking the localization of that biomolecule inside living...addition to emission wavelength range, there are other traits that need to be considered when choosing an FP:...chromophore. This can be from a few minutes after it is translated to a few hours. For example, superfolder GFP ...synaptic vesicles). Some FPs have different ex/em spectra (e.g. mKeima) or change fluorescent intensity upon...

...enhancers and genome math come in. They help us to transform raw genomic coordinates into meaningful experiments...of genome math. Figure 1: Genome math illustrated. Intersect (left) and set minus (right). ... Enhancers are regions of DNA that demonstrate “spooky action at a distance.” Through the marvel of DNA...“GPS for proteins”) for NANOG, an essential transcription factor in embryonic stem cells (ESCs). Your ...H3K4me1 + H3K27ac) – H3K4me3 The following video demonstrates how anyone can find enhancers with Quilt. ...and more. We know that this matter exists, but traditional approaches to study its function have been prohibitively...

...for Biology) on Reddit and got some substantial traffic that we thought it would be a good idea. After ...resources at her disposal, she was writing all day and straight on through her designated two hours for answering... general advice on how to meet people, how to transition out of academia, and other science career topics...diverse career path development--but the science trainees still have to lead on their own development. Please...resources. The internet content for scientists in training is abundant, helpful, and sometimes excellent....Sharing Your Lab Protocols: Using Apps to Save Time & Track Your Experiments Additional Resources See How ...

...need. Most often, early career scientists aren’t trained to have these types of negotiations. They usually... centrifuges, sequencers and analyzers, and any ultra sensitive equipment (RT-PCR, Microscopy, Fluorescence...list from your current P.I., lab manager, or administrator containing all of the materials that you’ve ...searches, and seek advice from your current administrative services, managers, and techs’ financial books...to proceed with a used or new item. To further illustrate this point, if your research is dependent on ...Channel your inner curiosity and use your research training to help you find other solutions to reach your...

...developments. These abrupt changes threaten to slow transformative biological discoveries and life-saving medical... can prove to be vital during this period of constrained funding. Addgene is uniquely positioned to help...archiving their biological materials during funding transitions. Enabling resource sharing: Our operating model...independently — particularly valuable when budgets are constrained. This collaborative approach eliminates redundant...Throughout history, the scientific community has demonstrated remarkable resilience and creativity in the ...

...characterized by single, linear, negative-sense, single-stranded RNA genomes. The virus does all of its damage ... also recently described how the glycoprotein transforms itself to escape vesicles within host cells and...critical component of vaccines and the target of neutralizing antibodies and inhibitors of attachment and ...California, Santa Cruz, offers sequence data on five strains of Ebola representing the latest outbreak and earlier...the current Ebola crisis, Science and Science Translational Medicine have made their complete collection...

...Gordon's lab developed a strategy to enhance HDR repair using a single-stranded oligodeoxynucleotide HDR...single-AAV delivery of Nme2Cas9 and the sgRNA to demonstrate efficient editing in adult mice and in zygotes...Shapiro’s lab. After testing dCas9 fusions with transcriptional repressors, they found that fusing dCas9 to...to an Mxi1 repressor domain repressed transcription of HSP90 an essential C. albicans gene. Read the ...

...-9 secretion, and identifying lysine methyltransferase substrates. We’re incredibly happy that lentiCRISPR... name suggests, lentiCRISPR v2 is a lentiviral transfer vector used to deliver both Cas9 and your gRNA...343.6166 (2014): 84-87. PubMed PMID: 24336571. PubMed Central PMCID: PMC4089965. 2. Sanjana, Neville E., Ophir...(2014): 783-784. PubMed PMID: 25075903. PubMed Central PMCID: PMC4486245. Resources on the Addgene Blog...

...complement our RNA Extraction Without A Kit blog post, we've compiled five methods for extracting DNA without...sharing even broader and began working with tech transfer offices to make reagents available to scientists...that cause plasmids to be incompatible. 4. RNA Extraction Without A Kit by Leah Schiesow We added to our...it, wrote these tips for working with RNA and extracting RNA without a kit. 3. SARS-CoV-2/COVID-19 Detection...

...containing the putative tracrRNA into a plasmid and expressed it in an E. coli bacteria strain. They found that...domain, characteristic of CRISPR-Cas enzymes. In contrast to other Cas enzymes, Cas14 has not been found...that Cas14 could indiscriminately cleave ssDNA in trans like Cas13 and Cas12 do for RNA and dsDNA, respectively...can cleave fluorescent probes of dsDNA and RNA in trans. Fluorescent signal indicates the presence of this...

...ability to direct enzymes – from nucleases to transcription factors – to specific sequences of DNA has become...and her colleagues led by Zhao described and demonstrated in a report published in The Journal of Biological...Biological Engineering in March is fused to a translational repressor. In principle, however, designer PUF...on a version that she hopes will change the intracellular localization of RNA. “You can think of changing...of designer PUF proteins for specific post-transcriptional regulation of endogenous RNA. J Biol Eng. 2014...

...GFP spreads through the cytosol via cytoplasmic strands. Image from Sattarzadeh et al., Sci Rep. 2015 CC...signal. Red signal intensity increases with GFP concentration and saturates around 30 uM. For low-abundance...most useful for labs that already have created a transgenic GFP-containing organism,” says Hanson, “using...unexpectedly photoconvertible GFP proteins to make a new transgenic line could avoid problems in localization that...Green to red photoconversion of GFP for protein tracking in vivo. Sci Rep. 2015 Jul 7;5:11771. doi: 10.1038...

...plasmid construct, most often in special E. coli strains designed for protein expression. Cultures are then...glycosylate proteins and may recapitulate the post-translational modifications added to proteins in other eukaryotes...the MISTIC (membrane-integrating sequence for translation of integral-membrane protein constructs) protein...(2014): e112643. PubMed PMID: 25386923. PubMed Central PMCID: PMC4227841. 2. Young, Carissa L., Zachary...Resources on the Addgene Blog Plasmids 101: E. coli strains for protein expression Plasmids 101: Protein tags...

...details how to transfect CRISPR plasmids into MEL cells by electroporation, sort your transfected cells via...genomic deletions We were inspired to produce intrachromosomal deletions based on the experiments of Kim ...CRISPR/Cas9, we began to explore the paired double-strand break (DSB) approach at a variety of loci. We were...Targeted deletions are just one of many available strategies by which biologists can imagine, create, and ...