...mouse and human kinomes. While any subset of the genome is limited by definition, the kinome is a well-annotated... About CRISPR Pooled Library Screening Why the kinome? While properly designed perturbations – in the...incremental approach is prudent. What's in the CRISPR kinome library? Thus, we are glad to provide CRISPR libraries...individual lentiGuide gRNA plasmids targeting the human kinome. These plasmids contain the same 3-4 gRNAs found... kinome...

...Image adapted from Ino et al. 2022, under CC-BY license.   Find MTRIA sensors here!   Ino D, et al. A fluorescent...develop a biosensor for oxytocin, Daisuke Ino and Hiroshi Hibino and colleagues devised a “Modular fluorescence...G-protein-coupled receptor (GPCR) recognizing a specific ligand (Ino et al. 2022). The oxytocin sensor MTRIAOT enabled...

...resource to have in your pocket.   5. +AMINO- by iGreenEarth AMINO is a very simple app that packs important...abbreviations of the amino acids memorized, but the structures are another story. AMINO has been very helpful...properties of or draw an amino acid or peptide for some reason. It contains all twenty amino acids plus selenocysteine...wishing that it had a codon list for each amino acid, but, overall AMINO can be beneficial for new researchers...important biological information, amino acid abbreviations, and structures into one convenient resource. ... Promega Material Safety Data Sheet by RSS-ICTD Amino Acid Structure Quiz/Card by Andrey Solovyev Lab ... He works with gene expression in Pseudomonas aeruginosa. Follow him on twitter @100percentSDS.     Additional...

...in which non-canonical amino acids (ncAAs, also sometimes called unnatural amino acids) are placed in a...re probably quite interested in the canonical 20 amino acids. But in your quest to learn more about your...protein of interest, you may find the available amino acids a bit…limiting. It may be time, then, to look...allowed for intentional incorporation of an unnatural amino acid in translation (Noren et al., 1989). In the...developed into an E. coli system, with the noncanonical amino acid in the media and the tRNAs and synthetase introduced...First, and most obviously, you need a non-canonical amino acid, which is usually added to the media for uptake...result in the incorporation of the non-canonical amino acid at the site of interest (figure 1). While the...

...genes encoding amino acid biosynthetic enzymes may be preferentially charged during amino acid starvation...preferences for the codons they use to encode specific amino acids. This is possible because there are 4 bases...There are therefore 64 possible codons but only 20 amino acids and 3 stop codons to encode leaving 41 codons...result is redundancy; multiple codons encode single amino acids. Evolutionary constraints have molded which... codon tables showing which codons encode which amino acids (see example to the right). With such simple...correctly, and charge the tRNAs with the appropriate amino acids. If an organism uses only a subset of codons...starvation thus resulting in higher production of amino acid biosynthetic enzymes (Dittmar et al., 2005). ...

...of as little as 8 or as many as several hundred amino acids, depending on the tag, and are introduced ...and cons. FLAG FLAG is a synthetically derived 8 amino acid tag (DYKDDDDK) that can be added multiple times...protein with enterokinase, which recognizes the 5 amino acids on the C terminal of the tag. Of the most ...with 5 negatively charged and 2 positively charged amino acids. FLAG tag has been used with moderate to high...as a chimera (rabbit). Myc The Myc tag is a 10 amino acid peptide (EQKLISEEDL) derived from the protooncogene...with 4 negatively charged and 1 positively charged amino acids. Myc tags are frequently used for western ...for all of these species. HA The HA tag is a 9 amino acid peptide (YPYDVPDYA) derived from human influenza...

...G418. Con: Slow transformation   Spectinomycin/streptomycin Spectinomycin and streptomycin both inhibit ...selection agent. Pros: Stable (spectinomycin). Cons: Cost (spectinomycin) or stability (streptomycin), ...Expensive.   Kanamycin Kanamycin, or kan, is an aminoglycoside which inhibits protein synthesis. The kan resistance...The NPTII gene confers resistance to several aminoglycosides, including G418 which is commonly used for ...carb (they are interchangeable). Like carb, spectinomycin is the more stable and expensive antibiotic ...antibiotic compared to streptomycin. Spectinomycin/streptomycin can be successfully used for plant and bacterial...purpose="decorative" title="Check icon" %} Spectinomycin Excellent No Just bacteria Not SHuffle cells...

...codons that encode 20 amino acids and three stop codons, meaning that the same amino acid can be encoded...actually prefer certain codons over others for certain amino acids. This is termed codon usage bias. In fact,...transfer RNAs (tRNAs). tRNAs bind to and deliver amino acids to the ribosome where they are incorporated...mRNA sequence. The 3’ end of  the tRNA binds the amino acid that corresponds to the anticodon sequence....always easy to identify the ideal codon for every amino acid across an entire polypeptide. Many researchers...wheel depicts the 64 codons that encodes the 20 amino acids and three stop codons. (B) Example of codon.... There are several codons that encode the same amino acid. Each organism, however, may preferentially...

...AraC system, which uses arabinose, requires many wash steps to remove arabinose making it difficult to ...🎶 “Bye bye L-arabinose drivePut your cultures in the shakerTurn the LEDs onAnd when you wantYou can ...recently published in @nchembio Title: Bye bye L-arabinose drive. Dedicated to Edo, Armin and @KhammashLab...lab,” says Di Ventura. It relies on the sugar arabinose to activate transcription by causing conformation...to regulate quickly. Arabinose also circulates within growth media and can’t be contained to a specific...AraC to make it responsive to light instead of arabinose. Nat Chem Biol. https://doi.org/10.1038/s41589...

...autoinflammatory loop: keratinocytes both respond to and release IL-36 cytokines. Keratinocytes are the main source...cytokine family resulting in the activation of keratinocytes, which release inflammatory mediators that trigger...known how IL-36 cytokines actually get out of keratinocytes and induce their effector functions. It’s an...important to study what is happening within the keratinocytes. “We hope to use CRISPR libraries from Addgene...or proteins are involved in IL-36 release in keratinocytes,” says Mellett. This would help the lab identify...

...for aminoacyl tRNA synthetases By Kate Harten Recombinant antibodies targeting fourteen aminoacyl tRNA...escape in SARS-CoV-2 Recombinant antibodies for aminoacyl tRNA synthetases Nanocompartments for yeast bioproduction...pooled plasmid library encompassing all possible amino acid mutations of the receptor binding domain (RBD... work by generating antibodies for additional aminoacyl tRNA synthetases.     Figure 2: Specific...encapsulated a rate-limiting enzyme in this pathway (myo-inositol oxygenase), providing enzyme protection while ...

...component of the nucleoside inosine. Created with BioRender.com. Inosine excision leads to the first...it to act on newly-deaminated inosines. Pro tip! The nucleoside inosine consists of the nucleobase hypoxanthine...repair. But mammals also have an enzyme to remove inosine from DNA: N-methylpurine DNA glycosylase (MPG),...extends to thymine Since an enzyme that excises inosine can be modified to target guanosine instead, perhaps...

...for longer. Cultures induced with arabinose and colonies on arabinose-containing plates will be visibly... inducible expression systems in E. coli. The arabinose-inducible promoter pBAD and a low-copy number ...wide range of inducer concentrations (0.1-1% L-arabinose) without signs of toxicity or burden relative ...or on LB-agar plates containing glucose and L-arabinose at 37oC for at least 12 hours, or at lower temperatures...after the E. coli have colonized tumors, inject L-arabinose intraperitoneally and wait 24 hours before imaging...

...1350 amino acids in size, which includes SpCas9, and shortern orthologs approximately 1000 amino acids..., only Staphylococcus aureus Cas9 (SaCas9, 1053 amino acids) displayed cleavage activity in mammalian ...that’s small enough to package into AAV. At 984 amino acids in length, Cas9 from Campylobacter jejuni ... Neisseria meningitidis Cas9 (NmeCas9) at 1,082 amino acids, can also be packaged into AAV. NmeCas9 has...these Cas proteins with AAV, at just 1,108 and 986 amino acids, respectively, the size of both is within ...

...Marketing Manager at sciNote. Tea Pavlek is a Product Marketing Manager at SciNote, interested in combining...research with tech advances. You an follow SciNote on Twitter @scinoteELN Additional Resources on the Addgene...blogger, Tea Pavlek, Product Marketing Manager at sciNote. Today, every lab has its own habits and approaches...priorities for future ELN development and developed the SciNote Electronic Lab Notebook. Implementing an electronic...

... Balla. "Live cell imaging of phosphoinositides with expressed inositide binding protein domains." Methods...be downsized by deleting residues. Only about 10 amino acids can be deleted from the C-terminus and about... but the development of non-natural fluorescent amino acids and their incorporation into proteins are ...homology domains can be used to detect specific phosphoinositides (Varnai and Balla, 2008) (Figure 2). RNA production...10. Hilaire, Mary Rose, et al. "Blue fluorescent amino acid for biological spectroscopy and microscopy....

...critical amino acid). Many common E. coli strains used for cloning are auxotrophic for specific amino acids...bacteria (one that doesn’t synthesize an essential amino acid), and a plasmid that complements the auxotrophy...acids. When plated on media lacking the essential amino acid, the bacteria will only grow if it acquired...containing the gene necessary to synthesize the missing amino acid. Auxotrophic selection is often used for cloning...

...mis-translation Bactericidal 50-100 ug/mL Spectinomycin aminoglycoside Binds 30S ribosomal subunit; interrupts...Action*   Working Concentration** Kanamycin aminoglycoside Binds 30S ribosomal subunit; causes mis-translation... Bactericidal 7.5-50 ug/mL Streptomycin aminoglycoside Inhibits initiation of protein synthesis ...macrolide Blocks 50S ribosomal subunit; inhibits aminoacyl translocation Bacteriostatic 50-100 ug/mL in...

...result in a spectinomycin resistant phenotype in bacteria. While there are many known spectinomycin resistance...increase the mutation rate of EvolvR, three additional amino acid changes were introduced to nCas9. These changes...five dispersed regions of the target gene of spectinomycin, rpsE. These results not only uncovered novel...used to generate bacteria resistant to both spectinomycin and streptomycin, but could also be used for...

...Acinetobacter calcoaceticus, Actinobacillus pleuropneumoniae, Actinomyces naeslundii, A. viscosus, Aerobacter...Providencia spp., Pseudomonas spp., Rhizobium leguminosarum, R. meliloti, Rhodobacter sphaeroides, R. capsulatus...Providencia stuartii, Pseudomonas spp., Rhizobium leguminosarum, R. trifolii, Salmonella enteritidis, S. typhimurium...fluorescens, P. putida, Rhizobium meliloti, R. leguminosarum by. viciae, Rhodobacter sphaeroides, Salmonella...