... brain where many different cell types, or even different phenotypes of the same cell type, are intermingled...: Flow chart depicting a) steps for selecting serotypes and promoters to test in your AAV system and b... b) steps for testing and optimizing your serotypes, titer, and promoter. Note that several steps in testing...are effected by multiple AAV variables.    Serotypes Serotype refers to the different protein compositions...determine how a virus enters the cell. Different serotypes utilize a number of different receptors, co-receptors...usually recommend testing more than one.  Most AAV serotypes have broad tropism (the ability of a virus to ...your AAV.  Figure 2: Comparison of AAV serotypes 1, 2, 5, 6, 8, and 9 in two different brain regions...

...AAV Serotypes and their target tissues (adapted from Addgene's AAV guide)  Commonly used serotypes include...untreated (for review see Dietzschold et al., 2008). Herpes Simplex Virus (HSV) also migrates through neurons...and VP3. To date several hundred variants, or serotypes, have been found naturally or have been genetically... animals that express Cre in only specific cell types, for example only dopaminergic neurons, neuronal...include AAV1 – 9, with most of these serotypes capable of entering neurons (see table 1)(Choi et al., 2005...anterograde intracellular transport of some AAV serotypes, but overall with rather low efficiency. Not all...Castle et al., 2014). Differences between AAV serotypes may be also linked to differences in viral uptake...

...only enters and delivers it cargo to certain cell types.  Botond Roska’s lab at the Institute of Molecular...existing promoters express in a broad number of cell types, such as inhibitory neurons, rods, cones, or retinal...that were transcriptionally active in retinal cell types in mice. Figure 1: The four strategies used...and 14% had targeted expression in specific cell types (ex: rods or cones) or cell classes (ex: photoreceptors...restricting AAV transgene expression to particular cell types or classes, these synthetic promoters have a few... few synthetic promoters targeted specific cell types, pairs of synthetic promoter can be used to restrict...a single cell type based on the overlap in cell types both promoters expressed in. The lab tested this...

...Universal. Can generate all adult cell types. Limited. Only specific cell types can be produced Ethical concern... Press reviews journal. Stem cells are special types of cells that can develop or “differentiate” into...cells’ ability to differentiate into multiple cell types makes them useful models for developmental processes...capability to differentiate into specialized cell types [2]. We general speak of 4 levels of stem cell ...including the embryonic and extraembryonic cell types. In mammalian development, cells found in the zygote...When a cell can differentiate into multiple cell types of the same lineage. Blood (hematopoietic) stem ... cells Stem cells are classified into two broad types according to their source: Embryonic stem cells...

...specific cell and tissue types. The combination of serotypes and psuedotypes means you can customize your...preference to infect specific cell or tissue types. Using serotypes that have a known tropism for your cells...BioRender.com. Beyond the identified serotypes, AAV also has psuedotypes. Psuedotyping is the process of making...capsids. To date, at least 12 different capsids, or serotypes, have been identified in nature, while many more...Figure 2: Visual representation of common AAV serotypes used for different tissues. Created with BioRender.com...capsid of another. Researchers have used these psuedotypes to further improve the tropism of AAV vectors...library to a significant degree using different types of AAV. Expression Another advantage of AAV vectors...

...are each suited to different types of modern day fluorescence microscopes (see Table 1). Thin dynamic ...Therefore these are sometimes call Epi-fluorescence microscopes. (See left image above for an example image)....by pixel is slow; therefore, certain confocal microscopes use parallelization to enhance performance. In...and a reduction in signal intensity. 2-Photon microscopes restrict excitation (and bleaching) to a single... scanning as is done in confocal or 2-photon microscopes. Total Internal Reflection Microscopy (TIRF) ...most difficult characteristic to capture in these types of samples is their fast movements. Depending on... with an evanescent energy field. Lightsheet microscopes that project ultra-thin sheets of excitation ...

... after the isotypes they interact with, so IgG isotypes bind to Fcγ receptors; IgE isotypes bind to Fcε...are part of the reason why different isotypes drive different types of immune responses. Handily, the receptors...that the Fc domain provides structure, determines isotypes, and provides a place for secondary antibodies...antibodies in our Antibodies 101 animation.)  Many types of immune cells, from mast cells to B cells, express...cells or responses. And there are still antibody isotypes and Fc receptors that we don’t know much about...Introduction to AntibodiesAntibodies 101: IsotypesAntibodies 101: Chimeric Antibodies  ...

... the research setting, antibodies of different isotypes can be used together in the same experiment because...small portion of the protein, known as an epitope. Epitopes are between 5-8 amino acids long, with a typical...are much larger than this, it may contain many epitopes that antibodies can recognize. Imagine you’re ... amino acids, or only in their 3D conformation. Types of antibodies and production In the immune system...extracted. These antibodies recognize a variety of epitopes on the target protein, so they are referred to...because the antibodies will recognize multiple epitopes on the protein. However, each animal will make...make antibodies against different protein epitopes and so variability between lots is a big concern. You...

...necessarily need to know where the epitopes are, just if the available epitopes could be affected by, or critical...change configurations or binding partners, the epitopes available for binding change as well. If this ...think about induced conformation changes. Several types of assays, such as ELISAs and Westerns, frequently...in an SDS-page gel. As the protein unfolds, new epitopes may appear, while others may get hidden. Monoclonal...well, don’t relax just yet. Proteins often change shapes and epitope accessibility naturally, by binding...only be recognized by antibodies that bind to the epitopes that are available on the outside of the cell....cell. The same protein may have different epitopes available if it is being selected out of a complex mix...

...for both in vitro and in vivo work with many cell types, as CAG is a ubiquitous and constitutive promoter...Optogenetics Yizhar New viral service with multiple serotypes pAAV_EF1a-DIO-PdCO-mScarlet-ER-miniWPRE AAV1...Optogenetics Yizhar New viral service with multiple serotypes pAAV_hSyn-PdCO-EGFP-WPRE AAV1, AAV5 Optogenetics...Optogenetics Yizhar New viral service with multiple serotypes pAAV-CaMKIIa(0.4)-eOPN3-mScarlet-WPRE AAV1, ...Optogenetics Yizhar New viral service with multiple serotypes pAAV-CAG-FLEX-rc [Jaws-KGC-tdTomato-ER2] AAV5...AAV5, AAV8 Optogenetics Boyden New serotypes pAAV-CAG-tTA AAV2 Molecular Tools Gradinaru New viral service...Biosensors Li New viral service with multiple serotypes pAAV-hSyn-GRAB_OTmut AAV9 Biosensors (Control...

...which maintains structural epitopes (orange region), but it can block epitopes hidden within the folded ... structure and thus structural epitopes, but can reveal linear epitopes previously hidden within the protein...these goals, researchers commonly use one of two types of fixatives - chemical cross-linkers and organic...components. This method of fixation is good for epitopes that are sensitive to cross-linking, but it doesn...while organic solvents can reveal otherwise hidden epitopes. However, keep in mind that you will still contend...the cell where they could bind to intracellular epitopes. C) Permeabilizing with a non-ionic detergent,...choose?! Ok, so now you know some of the possible types of fixatives or permeabilizers, but how do you decide...

... the selectivity of antibodies for other sample types and for non-surface (intracellular) antigens. Unfortunately...Zenodo community.  Knockdown approach For cell types where knockout studies are not feasible, an alternative...RNA or Proteomic data from multiple cell lines/ types with different expression Flow cytometry is often... to compare antibody labeling of different cell types within a sample to expected labeling (Figure 2).... allow for the clear identification of the cell types of interest. By correlating your data sets, you ...). If two antibodies, that recognize different epitopes of the same protein, show the same pattern of ...available to confirm that they recognize different epitopes. Detection of overexpressed (tagged or untagged...

... predict T cell epitopes focus on characterizing those epitopes as CD8 or CD4 epitopes, as well as their...the drug epitopes likely to be presented by T cells to elicit an immune response. T cell epitopes are found...are commonly used for the prediction of T cell epitopes based on the amino acid sequence of the antibody... assessment can help in determining the T-cell epitopes that will likely bring about unwanted immunogenicity...Monoclonal Antibodies and the Potential Use of HLA Haplotypes to Predict Vulnerable Patients. Frontiers in ...

...used with conventional microscopes, such as epifluorescence or confocal microscopes, but they can also be...long-term expression of AAV delivered genes. AAV serotypes have been extensively studied in order to define...Research Campus in the USA have used AAV1 and AAV2 serotypes in order to create an atlas of the mouse brain...enables researchers to quickly find the best AAV serotypes to target the neurons they’re studying. Thanks...detection when the POI is weakly expressed. As these epitopes tags have no intrinsic visual signal, they cannot...be expressed stably in neurons. As compared to epitopes tags, FPs are intrinsically fluorescent and can...using antibodies. In contrast, with their many epitopes, smFPs retain their ability to be detected after...

...most requested serotypes (Figure 4), our catalog includes vectors in a variety of serotypes, including the...be to use. AAVs, which can target different cell types depending on their serotype, are particularly challenging...in Addgene’s catalog are offered in different serotypes. In fact, the 879 viral vectors in our repository...Cumulative AAV preps for the seven most popular serotypes available from Addgene from 2016–2023. B) AAV ...AAV preps for the seven most popular serotypes distributed per year by Addgene from 2016–2023. How to... where we dove fairly deeply into the different types of tools scientists were using/depositing. Our viral...collaborations, and will continue to do so, in the hopes that these findings can help others as much as they've...

...from more cycles of proliferation Less Cell types All types Less Ease of genetic correction Easy Difficult...potential to produce all cell types in the body. On the other hand, only few cell types have been currently transdifferentiated...iPSCs have now been generated from many different types of somatic cells. The goal is to use cells that ... lineages [13]. Over the last decade, many cell types have been successfully differentiated from human...human iPSCs. Below is a list of some of these cell types [13]:  Germ cells [14] Hepatocytes [15] Pancreatic...for in vivo transdifferentiation. Multiple cell types such as fibroblasts, hepatocytes, and pancreatic...transdifferentiation is suited for switching cell types between similar lineages, reprogramming is more ...

...recombinase or a fluorescent marker in the various cell types of the basal ganglia. They work via newly-characterized...Optogenetics Ofer Yizhar New viral prep with multiple serotypes pAAV-Syn-FLEX-rc[ChrimsonR-tdTomato] AAV1 ... GENIE Project New viral prep with multiple serotypes pGP-AAV-GFAP-iGABASnFR2(no bind)-WPRE AAV1... GENIE Project New viral prep with multiple serotypes pGP-AAV-syn-iGABASnFR2-WPRE AAV1, AAV5 Biosensors... GENIE Project New viral prep with multiple serotypes pGP-AAV-syn-iGABASnFR2(no bind)-WPRE AAV1,... GENIE Project New viral prep with multiple serotypes pGP-AAV-syn-flex-iGABASnFR2-WPRE AAV1, AAV5... GENIE Project New viral prep with multiple serotypes Looking for more viral vectors? Browse the...

...system make different isotypes of antibodies that recognize many different epitopes of the target protein...polyclonal antibodies, one of the many different types of antibodies available (others you’ll encounter...protein G each have different affinities for IgG subtypes from different animals so it’s important to choose...due to their ability to recognize many different epitopes of the target protein. While this can increase... ability of polyclonal antibodies to bind many epitopes of their target protein thereby greatly increasing... antibody population recognizes many different epitopes, increasing the likelihood of cross-reactivity...

...mentors (super-agreeable friendly types, indecisives, and super busy types). If you supervisor ignores you...Coping with Difficult People,” identified seven types of difficult people: Hostile-aggressives, who are... and your questions, Super-agreeable friendly types, who don’t follow through on their promises, Wet-blanket...can bring down the morale of a group with their pessimism, Know-it all experts, who think their way is...of difficult supervisors: Hands-off super busy types, who never have time to mentor you, and Micromanagers...strategies to deal with each of these personality types is beyond the scope of this article, there are a...

...can also be directed to specific types of cells, such as neuron subtypes, thanks to viral serotype specificity...molecule which we will describe in more detail below. Types of Voltage Indicators GEVIs can be divided up structurally... BY 4.0.   FRET Chemigenetic Sensors These types of sensors can have a variety of different components... both. Targeting these systems to specific cell types is where viral vectors become invaluable. Viral...infection. Packaging viruses also have various serotypes – the surface proteins that confer affinity for... sensing!   Figure 3: Components of the types of voltage indicators reviewed.   References...