...References and Resources References Braun, C. J., Adames, A. C., Saur, D., & Rad, R. (2022). Tutorial: design... customization, and enhanced expression in many mouse tissues. Safety AAV and lentivirus have different...transduction efficiency over lentivirus in many mouse tissues. This will help improve your selection pool...screens have been used to map tumor suppressors in mouse liver, interrogate immune cells to improve immunotherapy...and generate programmable screening platforms for mouse tissues. Your AAVancement could be next!   References...Sousa, A. A., Tsai, S. Q., Bengtsson, N. E., Lööv, C., Ingelsson, M., Chamberlain, J. S., Corey, D. P.,...M. J., Joung, J. K., Breakefield, X. O., Maguire, C. A., & György, B. (2019). High levels of AAV vector...

...antibody, like if you are using an anti-mouse IgG antibody in mouse tissue. Such species-on-species staining...omitted a mouse IgG2a isotype primary antibody but you still observe a signal from your anti-Mouse IgG2a ... mice with the drug rather than rely on the results of a single mouse. Biological replicates help you ... slowing diseases progression in an Alzheimer’s mouse model, then you could treat one group of mice, the...increase the power of the results. In this case, mouse samples from positive (+), negative (-) and experimental...replicates such as multiple mice in each group and C, technical replicates or repeated measurements will...interactions between the sample and the secondary antibody. C, a label control treats the sample to the full protocol...

...the future of mouse work is bright as it is now easier than ever to manipulate the mouse genome using ... (for knock in models). CRISPR mouse model basics To create a mouse model, the gRNA, Cas9, and donor...of using CRISPR for mouse genome engineering From the very start, generating mouse models using CRISPR...fertilized mouse eggs, circumventing the need for enrichment and selection offered by mouse ESCs. For ...Generating Mouse Models Using CRISPR‐Cas9‐Mediated Genome Editing. Current Protocols in Mouse Biology 6... Haoyi Wang. CRISPR/Cas9 is revolutionizing the mouse gene-targeting field. Mice have long been extremely...extremely useful in the lab – they are relatively small and easy to work with, making them the go-to choice...

...FLAG antibodies on Addgene’s catalog including a mouse monoclonal (M2) as well as a chimera (rabbit). ...most prevalent clone used today! Goat, chicken, mouse, or rabbit? Addgene has you covered with monoclonal...sequence. Check out Addgene’s monoclonal antibodies (mouse, rabbit, and chicken) to this tag! GFP  GFP is more...Epitope tags are short peptides introduced at the N or C terminus of a protein that are bound by antibodies...enterokinase, which recognizes the 5 amino acids on the C terminal of the tag. Of the most common tags, FLAG...not at all. The introduction of a tag at the N or C terminus of a protein may disrupt the biological function...a specific application, or they exist but are unreliable for a variety of reasons (Baker, Nature). Tagging...

...R-mAb K58/35R (Mouse IgG2a) Goat anti-mouse IgG2a CASPR anti-CASPR mAb K65/35 (Mouse IgG1) Goat... Boeckman, J. X., Manning, C. F., Nguyen, J. T., Bechtold, H., Dumitras, C., Gong, B., Nguyen, K., Van..., you have Goat Anti-Rabbit IgG and Donkey Anti-Mouse IgG both with Alexa Fluor® 488) then you can’t use...Goat anti-mouse IgG1 Kv2.1 channel anti-Kv2.1 KC (Rabbit IgG polyclonal) Goat anti-Rabbit IgG ...-GABA-AR β1 (white) and anti-GABA-AR β3 (green). (C) Hippocampus labeled with anti-Kv2.1 (magenta), anti-AnkyrinG... Panels C1-C3 show magnified detail of the box in C. Image adapted from Andrews et al., 2019 under CC-... J., Kandov, E., Lowekamp, B., Speranza, E., Chu, C. J., Gola, A., Thakur, N., Shih, R., Yao, L., Yaniv...

...using a mouse model for a human neurological disease (Sinnamon et al., 2020). They introduced a mouse MECP2...10.1126/science.1206593 Powers S, Miranda C, Dennys-Rivers C, Huffenberger A, Braun L, Rinaldi F, Wein...al., 2007). They accomplished this using Cre-Lox mouse models in which the endogenous MECP2 gene was disrupted... ADAR2, into the hippocampus of a Rett Syndrome mouse model using an adeno-associated viral (AAV) vector... A (2007) Reversal of Neurological Defects in a Mouse Model of Rett Syndrome. Science 315:1143–1147 . ....2020.107878 Sinnett SE, Hector RD, Gadalla KKE, Heindel C, Chen D, Zaric V, Bailey MES, Cobb SR, Gray SJ (2017... expand as Rett scientists deposit their MECP2-related plasmid tools with Addgene and share additional...

...before working with mouse models, several common techniques used to create engineered mouse models at embryonic...understand mammalian traits and genetically engineered mouse models provide researchers with useful and adaptable...preclinical research. For scientists new to using mouse models, the possibilities may seem endless - and...editing techniques. Inbred and outbred strains Mouse strains used in research can fall under one of two...common genetic background, which means that each mouse is genetically identical and all mice of the strain... effect related to an experimental manipulation. Common inbred strains include C57BL/6, Balb/c, and 129...inserted at a specific site. To generate a transgenic mouse, microinjection of DNA or infection with a viral...

...should I pick? Do I go for the rabbit or the mouse? Is mouse IgG1 better or IgG2a? Will my choice affect... anti-mouse antibody. Others recognize specific isotopes within that species, like an anti-mouse IgG antibody... For example, an anti-mouse IgG2a secondary would be a better choice for a mouse monoclonal IgG2a than...species-on-species, i.e. using a mouse derived primary antibody to stain a target protein in mouse tissues, causes a ...are using mouse IgG2a and rat IgG1 antibodies in the same application, using a Goat anti-mouse IgG2a cross-adsorbed...antibody. Some are subclass specific, like an anti-mouse IgG2a antibody. When using a monoclonal antibody...than a general anti-mouse IgG. In a broadly specific secondary antibody only a fraction of the antibody ...

...small molecules targeting SMN1 and its partners in mouse cell lines. Dr. Di Schiavi hopes that these studies...mesothelioma that affects the lining of the lungs is relatively well known, there are a variety of lesser known...disease are unclear. Dr. Di Schiavi plans on using C. elegans as a model to identify SMN1 interacting partners...effects of smn1 disruption. As part of his work in C. elegans, Dr. Di Schiavi will use Addgene plasmids...these genes, CRISPR to disrupt their function, and C. elegans overexpression plasmids to upregulate their...research funding opportunities. References 1. Jones, C., et al. "PP09. 1–2352: Systematic review of incidence...heart health. These are amazing advances that will surely save thousands if not millions of lives, but many...

...Zhi Ying Chen, Robert C. Robbins, Mark A. Kay, Michael T. Longaker, and Joseph C. Wu. "A nonviral minicircle... generating iPSCs. The generation of iPSCs is relatively simple in concept: ectopically express a cocktail.... The advantages of this method are that it’s relatively simple to implement and doesn’t require time-... mesoderm, endoderm). Measuring pluripotency of mouse cells typically involves making chimeras by injecting...functional germ cells. The highest stringency test for mouse iPSCs, tetraploid complementation, involves injecting...measuring their ability to direct the development of a mouse. Since many of these functional tests aren’t possible...Generation of transgene-free induced pluripotent mouse stem cells by the piggyBac transposon." Nature Methods...

...Michael, S., Kuleshova, E., Ojala, D., Huang, C.-C., Gerfen, C. R., Schiller, J., Dudman, J. T., Hantman,...our in-stock viral prep service.  AAV9 AAV9 has relatively broad tissue tropism and high transduction efficiency...transport of AAV serotypes 1, 2, 5, 6, 8 and 9 in the mouse brain. PloS One, 8(9), e76310. https://doi.org/10.1371...Transfer Superior to AAV1, AAV6, AAV7, and AAV8 in the Mouse and Rat. Human Gene Therapy, 19(12), 1359–1368. ...doi.org/10.1089/hum.2008.123 Dalkara, D., Byrne, L. C., Klimczak, R. R., Visel, M., Yin, L., Merigan, W....scitranslmed.3005708 Goertsen, D., Goeden, N., Flytzanis, N. C., & Gradinaru, V. (2022). Targeting the lung epithelium...Ishikawa, Y., Jae, L. T., Wosen, J. E., Nagamine, C. M., Chapman, M. S., & Carette, J. E. (2016). An essential...

...is a recombinant version of the original V5 tag mouse monoclonal antibody, which recognizes a short, linear...applications.  IPI’s recombinant version is a fusion of mouse variable and rabbit constant domains and is recommended...Optimized mScarlet-based tags for live imaging in C. elegans CRISPR/Cas9 for gene disruption in M. abscessus...tagging proteins, as it can be fused to the N- or C-terminus (or even in the middle of proteins) and performs.... E., Young, D. F., Goswami, K. K., & Russell, W. C. (1987). Isolation and characterization of monoclonal...39406745. Optimizing mScarlet tags for live imaging in C. elegans By Alyssa Neuhaus Red fluorescent proteins...maturation rate limits its use in systems like the C. elegans embryo, where developmental processes happen...

... Sellnow, R. C., Boye, S. L., Coberly, B., Bennett, A., Agbandje-McKenna, M., Sortwell, C. E., Hauswirth...strains of the same species. What works well in a mouse may not work as well in a rat or non-human primate...Transport of AAV Serotypes 1, 2, 5, 6, 8 and 9 in the Mouse Brain. PLOS ONE, 8(9), e76310. https://doi.org/10.1371... vector serotypes 1, 2, 5, 8 and 9 in marmoset, mouse and macaque cerebral cortex. Neuroscience Research...Backer, M. W. A., Brans, M. A. D., Luijendijk, M. C., Garner, K. M., & Adan, R. A. H. (2010). Optimization...Dudek, A. M., Pillay, S., Puschnik, A. S., Nagamine, C. M., Cheng, F., Qiu, J., Carette, J. E., & Vandenberghe..., J. Y., Han, J. H., Lee, H., Han, Y. E., Rah, J. C., & Park, H. (2020). Labeling dual presynaptic inputs...

...transdifferentiation in vitro where treatment of mouse fibroblasts with 5-azacytidine led to their conversion...2000, Ferber et al. showed for the first time that mouse liver cells could be transdifferentiated in vivo...Yamanaka, Induction of pluripotent stem cells from mouse embryonic and adult fibroblast cultures by defined...PMC2758053. 8. Okita, K., et al., Generation of mouse induced pluripotent stem cells without viral vectors...four reprogramming factors- OCT4, SOX2, KLF4, and C-MYC - that induce a differentiated somatic cell to...additional pluripotency stage) Less Safety Less (C-MYC, a known oncogene is one of the reprogramming ...22645348. PubMed Central PMCID: PMC3390875. 25. Patsch, C., et al., Generation of vascular endothelial and smooth...

...window) Poulin JF, Caronia G, Hofer C, Cui Q, Helm B, Ramakrishnan C, Chan CS, Dombeck DA, Deisseroth K...Navarro M, Burnham N, Cristiano C, Dorrier CE, Tipton GJ, Ramakrishnan C, Kozicz T, Deisseroth K, Thiele...Markovic M, Wolff SB, Ramakrishnan C, Fenno L, Deisseroth K, Herry C, Arber S, Lüthi A. 2016. Midbrain ...including dual-transgenic recombinase-expressing mouse lines 2–9 and combinations of transgenic recombinase-expressing...window) of Flp-recombinase-expressing transgenic mouse and rat lines. The Fenno Lab maintains a public ... in order to enable three-feature cell targeting. C) Example of viral targeting of cells expressing Cre...processing, producing a functional molecular tool (C,F). Implementation The following resources may be ...

...with pLKO.1 C. Cloning shRNA oligos into pLKO.1 C.1 Recommended materials C.2 Annealing oligos C.3 Digesting...Virus may be stored at 4°C for a few days, but should be frozen at -20°C or -80°C for long-term storage...., R is a purine (A,G), and Y is a pyrimidine (C,U). G-C content should be 36-52%. Sense 3’ end should ...annealed oligo from step C.2 20 ng digested pLKO.1 TRC-cloning vector from step C.3 (If you were unable ...Store at 4°C. m. Add 5 mL of fresh media containing antibiotics to the cells and incubate at 37°C, 5% CO ...at 37°C, 5% CO 2 overnight. Day 2: b. The target cells should be approximately 80-90% confluent. c. Dilute...solution is stable at 4°C for up to one year. The powder form of polybrene is stable at 4°C for several years...

...Chain Fragment Variables (scFvs)Avoiding the Mouse on Mouse Mess in IHC Resources on Addgene.org  Browse...The modularity of antibodies means that it is relatively easy to physically divide up the different regions...fragments intended for use as primary antibodies are relatively uncommon. Though, if you want to use Fab fragments...features labeled. V = Variable domain. L = Light chain. C = Constant domain. H = Heavy chain. Created with biorender.com...

.... K., Chen, J., Pommier, G. C., Cogan, J. Z., Replogle, J. M., Adriaens, C., Ramadoss, G. N., Shi, Q.,...representative micrograph (C), particle orientation distribution and Fourier-shell correlation curve (D), and the... of delivery could be used to partially correct mouse models of genetic blindness (An et al., 2024) — ...could silence the expression of prion protein in mouse brains for at least six months. Unlike genome editing... module (SpyTag-GFP nanobody) assembled on beads. C) Micrograph of H1.8-GFP nucleosomes collected with...Protein samples protected from AWI by helical LEAs. C-E) cryo-EM single-particle analysis of PRC2 with RvLEAMshort...tardigrades!  Find RvLEAMshort here!    Abe, K. M., & Lim, C. J. (2024). Small LEA proteins as an effective air-water...

...-018-0219-4  Yu C-C (Jay), Barry NC, Wassie AT, et al (2020) Expansion microscopy of C. elegans. Elife...easier to expand than mouse brain slices, which are easier to expand than, say, a C. elegans (Damstra, et...physically separates molecules while maintaining their relative positions, allowing you to resolve individual ...accessible imaging equipment, ExM protocols utilize relatively inexpensive reagents and are readily adoptable.../doi.org/10.1038/nbt.3625  Truckenbrodt S, Sommer C, Rizzoli SO, Danzl JG (2019) A practical guide to ...they want to understand the ultrastructural relationships that influence everything from cell division...optimization and careful execution, but they cannot be entirely eliminated. It is recommended that you image samples...

...difficult to grow. Growth Temp Choose from 30 °C, 37 °C, or room temperature. Growth Strain Please... Choose from: H. sapiens (human), M. musculus (mouse), R. norvegicus (rat), G. gallus (chicken), B. taurus...expression, insect expression, plant expression, mouse targeting, CRISPR, TALEN, synthetic biology, affinity...frog), D. rerio (zebrafish), D. melanogaster (fly), C. elegans (nematode), S. cerevisiae (budding yeast)...be indicated in the “Species Other” column. Relevant Mutations Indicate the amino acid change as ...