... Zetsche et al., Nat Biotechnol. 2015 Feb 2;33(2):139-42. doi: 10.1038/nbt.3149. Light-inducible ...and on your favourite signalling pathways (Figure 2). They are all available now at Addgene. 1 Lungu...20;19(4):507-17. 10.1016/j.chembiol.2012.02.006. 2 Guntas et al., Proc Natl Acad Sci USA 2015 Jan 6; ...Nihongaki et al., Chemistry and Biology. 2015 Feb 19; 22(2):169-74. High efficiency cloning for enzyme screening...inducible, gene knockdown with shRNA or shRNA-miR30, (2) tamoxifen-inducible CreERT2 gene deletion, (3) simultaneous...different fluorescent proteins, 8 constitutive promoters, 2 includible promoters, 3 polyA terminators and various...of plasmids required for each input/output system; 2) removing all chemically inducible promoters; 3) improving...

...the system outlined in Schmid-Burgk, J.L., et al (2). In this post, I’ll cover two great applications ...molecularly evolved from mNeptune2 through 33 mutations and 2 deletions. CyOFP1 has a higher quantum yield, brightness...luciferin 24 hours later demonstrated a more than 2-fold brighter signal from Antares as compared to Fluc...PMID: 22894855. PubMed Central PMCID: PMC3501149. 2. Schmid, J.L., et al. (2016) CRISPaint allows modular...accuators. The researchers took a FRET reporter and adapted it into a BRET reporter by using NanoLuc® Luciferase...

...certain states of interest. For example, in Figure 2 below we present a two-input, five-state GRSM register...linking RSM inputs to endogenous gene expression (2). Our paper (1) explains how to arrange attB-attP...Science. 353, aad8559 (2016). PubMed PMID: 27463678. 2. Green, Alexander A., et al. "Toehold switches: de-novo-designed...straight-forward, concise explanation of how to adapt our engineered devices, recombinase-based state ...not expressed in each state. To help researchers adapt GRSMs for their research, we provide MATLAB-based...

...channelrhodopsin-2, commonly used in optogenetics, opening up the possibility of adapting GFP lines for ... Find plasmids from this publication at Addgene. 2. Tang, Jonathan C.Y., et al. “Cell type-specific manipulation...transcription activation domains. Both systems are easily adaptable to neuroscience applications, including optogenetics...

... PITCh vector via In-Fusion or SLIC cloning. Step 2: Design an insertion locus-specific gRNA The gRNA ...Find the plasmids from this publication at Addgene. 2. Dalvai, Mathieu, et al. (2015). “A Scalable Genome-Editing...method for genomic insertion. PITCh can also be adapted for whole-plasmid integration if you’d like to ...from a given promoter, and the technique can be adapted to other transgenes. It’s important to note that...increased versatility, it would be advantageous to adapt PITCh to insert genes into AAVS1, the “safe harbor...

...recognize Fab fragments from the same species and 2) these fragments will be more likely to bind (and ...epitopes recognized by the secondary.   Figure 2: There are a few common solutions to avoid background...-Mouse Immunohistochemical Staining Technique Adaptable to Biotin-Free Reagents, Immunofluorescence, and...

...activation is.   Figure 2: A bacteriograph of Michelangelo's "The Creation of Adam." This bacteriograph ...demonstrated through living images created by the team (Fig. 2). These images are based on the concept of bacteriographs...

...experiment using smFPs and standard neuronal tracers. 2. Visualization of neuronal sub-cellular structures...similar proteins. For example, N-cadherin (cadherin-2) is a postsynaptic cell adhesion protein that plays...PMID: 25915120. PubMed Central PMCID: PMC4573404. 2. Shepherd, G.M. (2004). In "The synaptic organization...smFPs in the future. References 1. Viswanathan, Sarada, et al. "High-performance probes for light and ...

... pCRISPomyces-2 contains Cas9 and a gRNA cassette. The simpler system of pCRISPomyces-2 displays a higher...and a subsequent curing protocol that requires only 2-3 hours incubation. E. coli and T. citrea scarless...four major components including (1) a CRISPR RNA, (2) four proteins forming the QCascade DNA-targeting ...functions more similarly to CRISPR/Cas9 systems adapted for eukaryotes in that Cas9-mediated cleavage induces...addition of a uracil DNA glycosylase inhibitor and degradation tag on the fusion protein to limit the activity...

...retain their individual activities when separated (2), and that DBD and TAs from different systems could... Enzymol. 1983;100:293-308. PubMed PMID: 6312261. 2. Keegan, L., Gill, G., and Ptashne, M. Separation ...folding, post-translational modification, protein degradation, access to the nucleus in eukaryotic screens ... system has been devised (8). References 1. Casadaban M.J., Martinez-Arias A., Shapira S.K., Chou J....Stynen B., Van Dijck P., Tournu H. A CUG codon adapted two-hybrid system for the pathogenic fungus Candida...

...due to the presence of additional helper sequences (2-3). As alluded to above, termination and polyadenylation...translation processes as well.   Figure 2: The conserved eukaryotic polyadenylation signal directs...longevity in eukaryotes and promote transcript degradation in prokaryotes.  Prokaryotic termination   Prokaryotic...important for stability of the mRNA, protection from degradation, and is integral to the nuclear export and translation...

...episode 2 "Organizing Literature" : I found out about Zotero here Dear Grad Student podcast episode 2 "Essential...This is a free web-based tool for detecting the readability of your writing. It makes suggestions on how ...

...the reporter mRNA is labeled by PCP-mCherry (Figure 2). The 3’ UTR also contains a CAAX sequence to bind...PMC2746483 Find ribosome profiling plasmids at Addgene. 2. Halstead, James M., et al. “Translation. An RNA biosensor...disrupting ribosome-mRNA complexes. Recent work has adapted this technique to monitor local translation and...

...host range plasmids. FEMS Microbiology Letters, 348(2), 87–96. https://doi.org/10.1111/1574-6968.12241  ...mobilization of broad host-range IncQ plasmids. Plasmid, 62(2), 57–70. https://doi.org/10.1016/j.plasmid.2009.05.001...negative hosts. Biotechnology and Bioengineering, 106(2), 326–332. https://doi.org/10.1002/bit.22695  Schmidhauser...contains structural elements that are versatile and adaptable. Variation in ori structure allows plasmids to...

...Indicators GEVIs can be divided up structurally into 2 main groups: voltage sensitive domains and Rhodopsin...depending on your experimental system.   Figure 2: General overview of chemigenetic tools. A genetically...the kinetic response time from signal input to readable output, which generally exceeds that of the VSD...of the previously described systems. Several adaptations of this method have been to tag, cage, uncage...

...that the signal to noise ratio can be increased from 2-fold (by CRISPRainbow) to 3-fold with CRISPR-Sirius...CNV) analysis of the U2OS cells.   Figure 2: CRISPR-Sirius tricolor labeling. (A) Diagram of sgRNA-Sirius...without changing them. Thus, it has been widely adapted for a wide variety of applications: base editing...

...misplacing your results or losing that lab notebook from 2 years ago. The free subscription to Lab Archives allots...notebooks you’re saving trees! The Earth thanks you. 2. Gibco Cell Culture by Gibco Whenever I start to learn...student, I have relied more and more heavily on adapting my smartphone to help me with my research in terms...

...ITR, which can either be “flip” or “flop” (Figure 2). Additionally, the ITR contains a 4-nucleotide Rep...site for Rep proteins (Savvy et al. 2013). Figure 2: AAV2 wild type ITR in flip and flop orientation. ...that can make vector propagation and sequencing a headache. How do I maintain ITR integrity and prevent unwanted...

...to a specific area at the plasma membrane (Figure 2). Just express the plasma membrane-localized iLID ...cells will be ready for the spotlight.    Figure 2: Opto-RhoGEFs to control Rho GTPase activity. A) Schematic...Schematic of Opto-katanin’s use in cells. Images adapted from Meiring et al., 2022, under CC-BY 4.0 license...blue light illumination. Scale bar 25 µm. Images adapted from Mahlandt et al., 2023, under CC-BY 4.0 license...vesicle trafficking via CRY2-CIB1 clustering. Image adapted from Aljiboury et al., 2023, under CC-BY 4.0 license...colocalizing with SuperPLDx30. Scale bar 10 µm. Images adapted from Tei et al. 2022, bioRxiv, under a CC-BY-NC-ND...doi.org/10.1371/journal.pgen.1010765.  Berlew, E. E., Yamada, K., Kuznetsov, I. A., Rand, E. A., Ochs, C. C....

...while increasing the standard mutation rate by only ~2-fold over nCas9 levels. Two sets of modifications ...10-3 mutations per nucleotide per generation, and (2) the addition of the thioredoxin-binding domain (TBD...Nature 560.7717 (2018): 248. PubMed PMID: 30069054. Sadanand, Saheli. "EvolvR-ing to targeted mutagenesis."...