...cells; function is unknown Monomer with a valency of 2 IgE Protects against parasites and is responsible ...such as anaphylactic shock Monomer with a valency of 2 IgG The most abundant isotype in blood, also found...secondary immune response Monomer with a valency of 2 IgG1; IgG2; IgG3; IgG4 IgM Produced during the primary... ranging from antibody fragments (shown in Figure 2) to completely unrelated molecules. Examples include...used for assembling protein nanostructures. Figure 2: An antibody (IgG, far left) compared to alternative... is described in more detail in the Multiplex Assays section below. The following table describes the ... Science Guides Antibody Guide Antibody Guide View Addgene's Antibody...

...196208 GB1-A11(2-196)-Strep ANXA11 His, Strep, Tev T7 ALS Lalit Deshmukh 196209 GB1-A11(2-196, G38R)-Strep...196211 GB1-A11(2-196, G175R)-Strep ANXA11 His, Strep, Tev T7 ALS Lalit Deshmukh 196212 GB1-A11(2-196, G189E...8661 p4455 FLAG-hPLIC-2 UBQLN2 Flag CMV ALS Peter Howley 8662 p4456 FLAG-hPLIC-2 NTF UBQLN2 Flag CMV ALS...Pietro De Camilli 194005 VPS13D1-1576-(Y1253S) VPS13D GFP CMV Spinocerebellar Ataxia Pietro De Camilli... Lalit Deshmukh 196210 GB1-A11(2-196, D40G)-Strep ANXA11 His, Strep, Tev T7 ALS Lalit Deshmukh 196211 ... ALS Lalit Deshmukh 196213 GB1-A11(2-52)-Strep ANXA11 His, Strep, Tev T7 ALS Lalit Deshmukh 196214 GB1... ALS Lalit Deshmukh 196216 GB1-A11(2-68)-Strep ANXA11 His, Strep, Tev T7 ALS Lalit Deshmukh 196694 mBFP-APP-mGFP...

...silencing, and desynchronization of neural activity, with single-spike temporal resolution. PLoS One. 2(3):e299...influx into the cell when stimulated. Channelrhodopsin-2 (ChR2), the first widely adopted optogenetic tool,... (Phy) interaction 660 Cryptochromes Cryptochrome 2 and CIB1 Blue-light–mediated induction of protein ...interactions based on Arabidopsis thaliana cryptochrome 2 and CIB1 450 LARIAT Inhibits protein function by reversibly...customizable TALE DNA-binding domain with cryptochrome 2 protein and its interacting partner CIB1 to control...transcription effectors OPTOSTIM PHR domain2 of cryptochrome 2 (Cry2) from Arabidopsis thaliana is fused to truncated...comparative analysis of microbial opsins. Nat Methods. 9(2):159-72. PMID 22179551 Saunders A, Johnson CA, Sabatini...

...the image can be acquired more quickly. 2-Photon Microscopy 2-Photon Microscopy attempts to solve two ...Fluorescence microscopy. Nature methods 2, 910-919 (2005). PubMed PMID: 16299476. 2. Conchello JA, Lichtman JW. Optical...enhance spatial contrast (i.e. optical sectioning (2) or super-resolution (3). Fluorescence microscopy...photobleaching and a reduction in signal intensity. 2-Photon microscopes restrict excitation (and bleaching...can be seen exiting through the tissue. Therefore, 2-photon allows for deeper imaging into thick tissues...imaging objective that collects fluorescence. Like 2-photon microscopy, only a single focal plane of the...relying on raster scanning as is done in confocal or 2-photon microscopes. Total Internal Reflection Microscopy...

...repository of tools for SAR-CoV-2 research. Two new antibodies, Anti-SARS-CoV-2 Nucleocapsid Protein [mBG17...Anti-SARS-CoV-2 Nucleocapsid Protein [mBG86], target distinct epitopes on the SARS-CoV-2 nucelocapsid ...Anti-SARS-CoV-2 Antibodies here! Terry, J.S., et al. (2021). Development of a SARS-CoV-2 nucleocapsid ...prime editor PE7 Antibodies to facilitate SARS-CoV-2 research New viral vector preps of Voltron2 voltage... incorporates La’s N-terminal domain, PE7 (Figure 2).  PE7 shows enhanced editing efficiencies in a number...editor. Find PE7 plasmids here!     Figure 2: PE7 improves prime editing. A) Schematic of PE7 prime...PMID: 38570691. Antibodies to facilitate SARS-CoV-2 research   By Ashley Waldron Ready-to-use recombinant...

...severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), the virus may acquire mutations to escape ... SARS-CoV-2 infects host cells through binding the human angiotensin converting enzyme 2 (ACE2) cell-surface...target of most anti-SARS-CoV-2 antibodies. In the library, each plasmid encodes the mutated spike protein...protein and a barcode, and is designed so that the SARS-CoV-2 spike RBD is displayed on the yeast cell’s surface...Pooled library to study antibody escape in SARS-CoV-2 Recombinant antibodies for aminoacyl tRNA synthetases...viral preps Mapping antibody escape in SARS-CoV-2 by mutational scanning By Seth Kasowitz As multiple...viral spike protein.   Figure 1. (A) SARS-CoV-2 RBD DMS Library generation and sequencing. From Starr...

...is an encyclopedia of Class 2 CRISPR systems with wiki entries describing enzyme activity, PAM requirements...Protocols gRNA Design Tools CRISPR Blog Posts The table below lists gRNA plasmids designed to target various...pyogenes , S. aureus etc.) was this gRNA sequence designed for? A given gRNA sequence may only be compatible...unpublished plasmids containing gRNA sequences designed to be used with any CRISPR nuclease and function...

... State University. Since its appearance, SARS-CoV-2 has spread to almost every part of the world manifesting...Rapid detection of 2019 novel coronavirus SARS-CoV-2 using a CRISPR based DETECTR assay Sensitivity:10-...primers that target the E and the N2 gene of SARS-CoV-2. If the Cas12a-sgRNA complex binds to the target sequences... method has been optimized to detect the SARS-CoV-2 virus directly using saliva without having to perform...and visual detection of novel coronavirus SARS-CoV-2 Sensitivity: 4.6-11 copies/microliter of RNA targets...single-molecule sensitive. In the case of HIV-1 and SARS-CoV-2 detection, AIOD-CRISPR without preamplification was...easy-to-deploy, protocol for Cas13-based detection of SARS-CoV-2 genetic material (CREST) Sensitivity: 10 copies of...

...gene-specific CRISPR and donor vectors as well as pX330S-2-PITCh, expressing a generic gRNA targeting the PITCh...Protocols, 2016 . Yamamoto Tagging Plasmids: pX330S-2-PITCh - expresses Cas9 & PITCh-gRNA pX330A-FBL/PITCh...Resources CRISPR Guide Viral Preps Protocols gRNA Design Tools CRISPR Blog Posts The CRISPR system has been...plasmids and protocols needed for using them, are described below. Read our CRISPR 101 post for more information...information about HDR/repair template design. How to use CRISPR to tag your gene of interest Mendenhall and...interest is currently unavailable, you will need to design and clone in a gRNA(s) to guide the Cas9 protein...protein to your target sequence, as well as design and clone in the homology arms for the donor plasmid. The...

...optogenetic tool is the widely used Channelrhodopsin-2 (ChR2) from Chlamydomonas reinhardtii . ChR2 is a ...many different types of proteins. For a full description (including peak response spectra) of different...preparations of many optogenetics plasmids. ID Plasmid Description Gene/Insert Vector Type Promoter PI Publication...transcription", "Hahn", or other terms. ID Plasmid Description Gene/Insert Vector Type Promoter PI Publication...

...AAV vectors for trial purposes in serotypes AAV1, 2, 5, 8, 9 and retrograde Retrograde AAV To target projection...comprehensive high-quality quality control process includes titering, purity assessment, sterility testing...

... discovery, 2, 952326. https://doi.org/10.3389/fddsv.2022.952326Joubert, M. K., Deshpande, M., Yang, J...various components of the immune system. Figure 2: In-vitro models for immunogenicity prediction of ...drug reaction. This system would somewhat act like a 2-step verification. All approaches have their own limitations...Terry, F. E., Roberts, B. J., Rosenberg, A. S., & De Groot, A. S. (2022). In silico Immunogenicity Assessment...in vivo animal models. In these cases, they are designed to mimic the human lymphatic system or even the...

...either SARS-CoV-2, Adenovirus type 41 (AdV-41), or Adenovirus-Associated Virus type 2 (AAV2) – or some...either 1) a physiological effect of SARS-CoV-2 toward destabilizing one’s natural immune response to Adenovirus...Adenovirus (and/or to its partner, AAV) or 2) a sociological effect of the COVID-19 lockdowns causing a lack...adenoviral vector infected cells does not require de novo viral gene expression: Implications for gene...… Chiu, C. Y. (2023). Adeno-associated virus type 2 in US children with acute severe hepatitis. Nature...viruses infecting humans (from seven species, designated as letters A–G) exist. The life cycle of AdVs...short-lived. It is worth noting that it is possible to design a vector that achieves persistent transgene expression...

...indicates a virus containing the genome of serotype 2 packaged in the capsid from serotype 5. Use of these...neurons that are not efficiently transduced by AAV2/2, and is distributed more widely in the brain, indicating... pseudotyped with viral capsids from serotypes 1, 2, and 5 display differential efficiency and cell tropism...PJ, Mandel RJ, Muzyczka N. Mol. Ther. 2004 Aug;10(2):302-17. PubMed . From virus evolution to vector revolution... Science Guides AAV Guide Adeno-associated Virus (AAV) Guide Components...Viral serotypes and pseudotyping/gene therapy: Designer gene delivery vectors: molecular engineering and...

...identification.   Fig. 2: Gel image of Plasmid 61564: pBAMD1-2 (~4.7 kb) digested with SpeI and... at ~3.6 kb and ~1.1 kb (lane 3). Uncut DNA (lane 2) has a band at <3 kb, which corresponds to the supercoiled...formation of plasmid multimers. Cell. 1976 Dec;9(4 PT 2):707-16. doi: 10.1016/0092-8674(76)90134-3. PMID: ...Strausbaugh LD, Gray P, Vartak NB, Brown M, Talbot D, Berg DE. pBR322-derived multicopy plasmids harboring large...dimers in Escherichia coli K-12. Plasmid. 1989 Mar;21(2):138-41. doi: 10.1016/0147-619x(89)90057-7. PMID: ...Plasmids and Chromosomes. Microbiol Spectr. 2014 Oct;2(5). doi: 10.1128/microbiolspec.PLAS-0025-2014. PMID...Behavior of Plasmid DNA. Microbiol Spectr. 2015 Apr;3(2):10.1128/microbiolspec.PLAS-0036-2014. doi: 10.1128...

...published an article on BioRxiv describing a comprehensive SARS-CoV-2 virus-human interaction network ...comprises all SARS-CoV-2 viral proteins except for Nsp3 and Nsp16, and includes an additional catalytically...all became acutely aware of the threat of SARS-CoV-2. Like many in the science community, we’ve been motivated...network in which we expressed 26 of the 29 SARS-CoV-2 viral proteins in human cells and used these viral “bait...identifying these potential human helpers of SARS-CoV-2 (especially the ones that are druggable) is a key ...just at the beginning of understanding how SARS-CoV-2 interacts with human proteins. For example, we used... joining in this worldwide fight against SARS-CoV-2. As we’ve all been affected in various ways by the...

... non-physiological, and difficult to ignore [1, 2]. 2. Fusion to an obligate dimer or oligomer Several...Biol, 2003. 163(2): p. 257-69. PubMed PMID: 14581454. PubMed Central PMCID: PMC2173526. 2. Zacharias, D....changes that the biosensor is designed to detect resulting in false positives [2, 4]. Solutions to the polymer...an A206K mutation [2]. Addgene has many of these constructs and they are designated mEGFP, mVenus, etc...analytical ultracentrifugation [2]. Some affinity assays used in the papers describing the FPs above including... nature are prone to dimerization (i.e. EGFP) [1, 2] or even form obligate tetramers (i.e. DsRed) [3]....form dimers in a cell. The Kd of EGFP is 0.11 mM [2]. Following the simple logic outlined above, you might...

...-CoV-2 receptor-binding domain. BioRxiv.  https://doi.org/10.1101/2020.04.16.045419.  Wrapp D, De Vlieger...select for optimal sybody candidates. Figure 2: Outline of the process for selection of sybodies ...identify and isolate sybodies targeting the SARS-CoV-2 Receptor Binding Domain found on the membrane-bound...Generation Toolbox kit. Find the Seeger lab's SARS-CoV-2 sybodies! References Geertsma ER (2013) FX Cloning...Zimmermann et al., 2020). This selection platform was designed so that any standard lab can quickly select for...strongly to your target protein. The Seeger lab designed 3 different mRNA sybody libraries for use in the... randomized with a defined set of different trinucleotides (corresponding to different amino acids) in...

...References Akhtar, W., De Jong, J., Pindyurin, A. V., Pagie, L., Meuleman, W., De Ridder, J., Berns, A....some of the best designs (e.g., top 5) and test them in their labs. Figure 2: Guidelines to predict... flanking sequences to their input string (Figure 2). This string can then be used as input for the prediction...Biotechnology. https://doi.org/10.1038/s41587-024-02268-2 Schep, R., Brinkman, E. K., Leemans, C., Vergara, ...this means we could design hundreds to thousands of different pegRNAs for each desired edit, it simply becomes...via GitHub (which includes prediction in batch mode). To predict the best pegRNA design for a specific edit...or offline via GitHub. The model provides users with different designs of pegRNAs, ranked by their PRIDICT...

...chromosomal integration of DNA. ACS Synthetic Biology, 2(9), 537–541. DOI: 10.1021/sb400021j Figure 1: Multiplexing...CRISPR guides. The first one binds to sfgfp with either 0, 10, 11, 14, or 20 matching nucleotides, the ...also like... CRISPR Guide CRISPR Protocols gRNA Design Tools CRISPR Blog Posts The David Bikard Lab at...mcherry with either 0, 11, 18, or 20 matching nucleotides. Together, these plasmids allow to explore the...