...IgM, IgD, and IgG are deleted.     Figure 2: Isotype switching requires DNA recombination of the...molecules are recruited by the antibody to help destroy and remove a pathogen. Different isotypes also ...well as the genes that encode those proteins are designated by the lower-case Greek letters μ, δ, γ, ε, and...

...available in the adenovirus section below!   Fig. 2 - Viral vector components of AAV Adenovirus Adenoviruses...promoter should flank this component as well. On both sides of this promoter-gene combo are two long terminal...Laboratory Press; 1997. Principles of Retroviral Vector Design. Available from: https://www.ncbi.nlm.nih.gov/books...

...higher quenching of mScarlet emission. Figure 2: Ace-mScarlet is a fusion between the voltage-sensitive...showing the potential for broad use. This kit is designed for use with acceptor vectors from the MoClo Toolkit...tracing Article contributed by Erin Sanders CreLite, designed by the Eisenhoffer lab, is a new tool that increases...

..., N. and Hamilton, D. 1981. PubMed PMID: 6276558. 2. Site-specific DNA recombination in mammalian cells...using a resistance marker; however, it is often desirable to remove the marker after the initial selection... Combining this with some of the loxP methods described above, a genetic modification can be restricted...

...1) or added using PCR-based amplification (Option 2). Advantages of Golden Gate cloning Golden Gate cloning...customized to direct assembly of DNA fragments. When designed correctly, the recognition sites do not appear... scheme is as follows: the gene of interest is designed with Type IIS sites (such as BsaI or BbsI), that...and do not appear in the final construct. The destination vector contains sites with complementary overhangs...ligation can be done in one 30-minute reaction. The destination vector and entry vector(s) are placed in a single...IIS enzyme and ligase. Although the original destination vector + insert may spontaneously religate, this... be re-digested. In contrast, formation of the desired ligation product is irreversible because this construct...

... later this year. Designing the donor plasmid Figure 2: Donor plasmid design strategy. A donor ...with each cell line. The editing strategy and design described above can be used to create similar donor ...GFP tag and 1kb of homologous DNA flanking both sides (about 2.7kb total)—is inserted into a plasmid backbone...plasmid to prevent the plasmid from being cut and destroyed by CRISPR/Cas9 during transfection. Making the...use electroporation, a technique that briefly destabilizes the cell’s outer membrane and allows the components...Addgene’s CRISPR-based Protein Tagging collection includes systems for tagging mammalian cell lines, Drosophila...

...still depolarize neurons at lower MOIs. Figure 2: ChRgers are delivered by intravenous injection into... model to help them construct “designer” channelrhodopsins with desired characteristics. The model was...Viviana Gradinaru’s lab used machine learning to design new channelrhodopsins that are more light sensitive...constrained and poorly understood. It’s hard to design new channelrhodopsins with high-throughput proteins...Using machine-learning to overcome channelrhodopsin design challenges To overcome these challenges, the Arnold...experiments. The sequences in this library were designed using structural information about parental channelrhodopsins...existing channelrhodopsins CoChR and/or ChR2(H134R). Designer channelrhodopsins paired with a systemic AAV allow...

...neuron-astrocyte, and cancer-stromal cell pairs.   Figure 2: Schematic of the interaction between the sender cell...repository that will help epigenetic researchers custom design their own engineering plasmid tools in about a ...cloning, the Haynes lab’s Protocols.io protocol describes an elegant system that allows for the assembly... cells that make up the human body and G-baToN provides versibility in tracking and modulating the interactions... inhibitor response 1 (TIR1) expressed under a desired promoter with a specific spatiotemporal expression...interests via CRISPR techniques. This tool-kit includes an array of FPs as well as additional useful tags...libraries encode target sites flanked by randomized nucleotides in place of a protospacer adjacent motif (PAM...

...Antibody Fragments. Antibodies (Basel). 2019 Apr 9;8(2):28. doi: 10.3390/antib8020028. PMID: 31544834; PMCID... as the OG (as the kids say) affinity reagents deserved to be celebrated, there are many other amazing...its thorns and every affinity reagent has its downsides. These reduced-in-size options often have reduced...°C and can refold correctly after denaturing.  Designed ankyrin repeat proteins, or DARPins, are genetically...a wide variety of targets, including proteins, peptides, amino acids, drugs, metal ions, and cells. They...

...Era of Personalized Medicine. Genes Dis. 2017 Jun;4(2):43-63. DOI: 10.1016/j.gendis.2017.04.001 Maetzig...about a specific virus? We have you covered with guides on lentivirus, retrovirus, adenovirus, and AAV....

...improvement in muscle physiological performance.  2. Trans-splicing Overview: For trans-plicing, splice...doi.org/10.1002/emmm.201302948 Trapani I, Toriello E, de Simone S, Colella P, Iodice C, Polishchuk EV, Sommella...physical size of the genomes is still ~4.7kb. But despite this, larger functional transgene products were...HDR event can’t occur. Lastly, depending on its design, donor A may be capable of expressing a truncated...

...signal separately for each target.   Figure 2: Examples of simultaneous multiplex IF labeling of... Brewer, M., McDonough, L., Zhu, Y., Neumann, E., De Caestecker, M., Gutierrez, D., & Spraggins, J. (2022...between the targets. Or, it might be sufficient to describe or classify the labeled structures visually; for...will vary depending on your specific experimental design. Like selecting an antibody, some of the requirements...

...inside the cell,” says Pedre Pérez. Figure 2: Micrographs of the biosensor expressed in the human...sulfurtransferase (MPST) which catalyzes the desulfuration of 3-mercaptopyruvate and generates an enzyme-bound...transferred to thiol-containing acceptors. These hydropersulfides modulate protein activity and have important...

...beginning Let’s take a quick flash back to 2004. “Shrek 2” was the most popular movie in theaters, the Razr ...business practices, like accepting e-signatures and designing a website users could easily navigate. This meant...Read the Addgene guide: https://www.addgene.org/guides/crispr/” — SirHamhands, responding to a request...

...main decisions to make: 1. Which modification? and 2. Which viral vector should be used? Modifying gene...behavioral neuroscience, and address experimental design considerations used in all behavioral studies. ...ion channels. Similarly, chemogenetic tools are designed channels that are chemically activated. Both are...

...strain (which will be covered in more detail in part 2 of this blog series) controls the cell types in which...reporter gene, which replaces the targeted gene and provides a way to track successfully engineered cells. ...genotype each animal to identify the ones with the desired genotype. Inducible and conditional systems Conditional...

...company culture.  It’s also a good idea to prepare 1-2 unique questions for each interviewer by looking at...method seems simple, I found it hard to remember to describe all four parts of a STAR answer once the interview... because sometimes companies have relaxed dress codes for interviews. You can also ask HR for clarification...

...protocols 4.4 (2009): 495-505. PubMed PMID: 19300443. 2. Geraerts, Martine, et al. "Comparison of lentiviral...virus will not only ensure that your infection is designed in the best possible way, but it may also save...the transgene itself. Many researchers prefer to design primers that target the common features of viral...

...communicate the nuances of protein structure in a flat 2-D images. In the early days of structural biology,...BioMolecular Modeling and the CEO of 3-D Molecular Designs. Have you ever held your favorite protein in the...favorite protein in your hand? At 3D Molecular Designs, we create physical models of proteins using 3D...off a small, family-run business – 3D Molecular Designs – to commercialize some of the best instructional...reaction to this model on his blog. Or, for a description of how effective this model was in explaining... up close and personal, visit the 3D Molecular Designs web site. Or, if you are visiting a local classroom...check out our hands-on teaching tools that are designed to make the molecular world real for students....

...unc- 119 + selection." MicroPublication 1 (2018): 2–3. Wang, Han, et al. "cGAL, a temperature-robust GAL4...of 3’UTRs, the Arizona State University (ASU) Biodesign institute developed a 3’UTR database specifically...specifically for C. elegans. This database provides detailed information about C. elegans 3’UTRs structures, ...imaging processing. The combination of these methods provides unique opportunities to develop computational ...Addgene In addition to the wealth of open resources described above, there were many plasmid tools presented...self-excise, and covalently ligate their flanking peptides together (Wang et al., 2018).” With the split ...continuously curated as we continue to receive plasmids designed for the worm so check back frequently for what...

... Cure for HIV/AIDS. Int J Mol Sci. 2023 Jan 13;24(2):1563. doi: 10.3390/ijms24021563 Resources on Addgene.org...introducing a desired edit, but this particular class of treatments has quite the opposite aim: to destroy. One...potential for treatment is no longer an accurate descriptor; progress in clinical therapy is more fitting...One treatment currently in clinical trials includes Cas3 (the shredder member of the Cas family) as part...CRISPR therapies have made significant clinical strides since we first wrote this blog post. The future...

...C-terminus of human codon-optimized  cryptochrome 2 (CRY2) from Arabidopsis thaliana. When exposed to ...soCoChR-expressing system. Taken together, Shemesh et. al. describe a novel system for the precise mapping of neuronal...the single-cell level. Various soCoChR plasmids described in this work including the soCoChR-GFP plasmid...information about spatial and temporal organization. The design use to detect PKA activity can be generalized to...choice for FRET assays where a dark acceptor is desired due to its very low quantum efficiency. ShadowY...

...156–159. https://doi.org/10.1016/0003-2697(87)90021-2 Chomczynski P, Sacchi N (2006) The single-step method...Allewell and Sarma, 1974). Additionally, ThermoFisher provides a protocol on how to integrate RNAlater® with ...

... adapted to a competitive format. Figure 2: Overview of using Competitive ELISA to quantitatively...ELISA is a modification of the sandwich ELISA as described here, the direct and indirect ELISA (more details...

... to remove this default setting. Timesaving Tip #2: blastx searches are inherently slower than blastn...terrific getting-started guide for BLAST, which includes a simple explanation of the different BLAST programs...matrix BLOSUM62, but you may want to check the description of the other matrices to see if another would...alternatives Depending on your sequencing result and desired analysis, BLAST may not always be your optimal ...

...more tips on using social media as a scientist here. 2) Public engagement - Participate in science festivals... Guardian, BBC Breakfast or the Londonist. BBC provides a peak into their day-to-day business with popular...Co-Ordinating Centre for Public Engagement (NCCPE) provides on-site courses in public engagement for institutions...

...non-specific binding in our IHC condition.   Fig. 2: Schematic of the goldilocks antibody selection process...perform differently over time), compatible with the desired applications (such as immunohistochemistry (IHC...antibodies, we have developed a criterion that includes accuracy, strength, and reproducibility. We test...

... Then he desribed how P. furiosus has systems to defend against both DNA and RNA viruses - 2 cas systems...plasmids, especially those of the CRISPR variety (see slides from my talk below). Largely organized by Dmitry...

... the Cas machinery where to cut.      Fig. 2: Heatmap indicating loci where SNPs reside on sgRNA... reference?  The CRISPR guides used in the screens they analyzed were designed to have a perfect match...African descent have approximately 20% more single nucleotide polymorphisms (SNPs) that map to guides in the...accounted for in the CRISPR guides they used, leading to mismatches between the guides and the lines’ genomes...modify the CRISpick algorithm to prevent guides from being designed that target regions with high genomic...coming through an unexpected source: the CRISPR guides, short pieces of RNA that bind to a cell’s DNA,...x-axis) which SNPs map to thetargeting sequences of guides across 8 CRISPR libraries. Samples are divided ...

...of these issues are not particularly new. Chapter 2, “The Professional Scientist,” addresses the challenges... Zimmer provides several. In Part Three of the book, called simply “Old Science,” he describes the now...teaching and communicating science for over two decades, attempts to answer it in five paragraphs, with...recent resurgence of “Do-It-Yourself Science” described in Chapter 3, and of course the rapidly changing... science.”  The three chapters in this section describe the challenges of trusting science in a “fake ...changed significantly over the last decade. He describes the detection of gravitational waves--a story ...collaboration from scientists around the world. The paper describing the waves’ detection had over 1,000 authors. ...