...IBEX is a highly multiplexed tissue imaging method that uses iterative staining and chemical bleaching...Radtke et al., 2020, 2022). I do mean highly multiplexed: you can image over 65 different parameters using...an invaluable resource for anyone performing multiplexed imaging, not just IBEX. The IBEX Imaging Community...., & Germain, R. N. (2020). IBEX: A versatile multiplex optical imaging approach for deep phenotyping ...resources on the Addgene blog Antibodies 101: Multiplex Immunofluorescence Negative Can Be Positive: Open...

...and also makes it hard to compare results across multiple targets because different CAPTURE cell lines will...lab presented three ways to use CAPTURE 2.0: 1. Multiplex capture of chromatin interactions The Xu lab performed...experiment to show that CAPTURE 2.0 could detect multiple chromatin interactions at once. Chromatin interactions...Long-range chromatin interactions identified with the multiplexed CAPTURE 2.0 largely replicated interactions previously... a large cluster of enhancers. CAPTURE 2.0’s multiplexing and high resolution allowed the Xu Lab to simultaneously...role in gene regulation The high resolution and multiplexed chromatin ‘photos’ generated with CAPTURE 2.0..., Orkin, S.H., Zhang, M.Q., & Xu, J. (2020). Multiplexed capture of spatial configuration and temporal...

...Stochastic multicolor labeling is a popular technique in neuroscience and developmental biology. This...or entire organism. Because each cell can have multiple copies of the transgene that will recombine independently...persistently fluorescent during development. Stochastic multicolor labeling systems, many based on Brainbow, now... all cell types can be labeled, and that while multiple copies of the transgene can integrate into cells...improving the level of detail possible for stochastic multicolor labeling of the brain. Labeling Drosophila neurons... of transgenic zebrafish lines that allow for multicolor labeling of an entire organism or a specific ...Zebrabow as a system that takes advantage of the multicolor labeling possible from Brainbow. Tracing clonal...

...MoClo application is to assemble multiple unique sgRNAs into a multi-TU targeting construct (Cas9, sgRNA...popular DNA assembly tool used to build single- and multi-gene constructs for a variety of applications. MoClo...favorite gene-of-interest, all the way to the complex multi-TU constructs that compose genetic circuits and ...bacteria MoClo is commonly used to design TUs and multi-TU constructs for synthetic biology applications...assembled into individual TUs or very large (up to 20!) multi-TU constructs (Moore et al., 2016). The variety ... 2016). CIDAR offers a one-pot reaction wherein multi-part assembly can occur. It is quite extensive with...interactions in gene expression, and to generate multi-gene knockouts. Mammalian and fly applications MoClo...

... construction system for CRISPR/Cas9-mediated multiplex genome engineering is now available thanks to ...new tool in Scientific Reports in June. “The multiplexity is one of the most advantageous properties of...established way of making an all-in-one vector for multiplex genome engineering.” Now, they’ve filled that... also be expanded to include even more loci. Multiplex genome engineering demonstrated in human cells...Cas9-nickase vectors work successfully to target multiple sites in cultured human cells. While it has not...be applied to efficient viral vector-mediated multiplex genome engineering. The system now provides an...an efficient targeting strategy for multiplex editing of genomes. It will also allow simultaneous activation...

...can these AAV’s be used for?  1. Multi-color labeling of neurons Multi-color labeling of neurons is for...fluorescent neurons. A simple but flawed approach to multi-color labeling is to inject a mix of AAV vectors...less cumbersome than brainbow, another method for multi-color labeling of neurons, because it doesn’t require... system that allows for controlled induction of multi-color labeling of neurons (figure 4). These capsids...lots of individual cells will be transduced with multiple viral particles and labeled a variety of colors...

...Find CROPseq-multi plasmids here Walton, R.T., Qin, Y., Blainey, P.C. (2024). CROPseq-multi: a versatile...developed a multiplexing approach to Cas9-based pooled CRISPR screens called CROPseq-multi (Walton et ...Figure 1: CROPseq-multi uses two sgRNAs with internal barcodes (iBARs), multiplexed using tRNAs, within...human cells. Addgene offers vectors for CROPseq-multi using selection by Puromycin, Zeocin®, or nuclear...articles.  Here's what you'll find in this post: Multiplexed perturbation and decoding in pooled CRISPR screens...System Neurodegeneration Collection updates   Multiplexed perturbation and decoding in pooled CRISPR screens...reduce recombination and avoid separation of the multiplexed elements. The team designed sgRNAs that include...

...complexed into multi-TU constructs, known as multigene constructs. Construction of multigene constructs, ...cloning, or MoClo, is an organization scheme for multi-part DNA assembly. Simply put, it assembles individual..., through multi-TU assembly,happens via three steps: Level 0: Cloning of basic parts (promoters, UTRs...useful, you may ask? You might be surprised by how multi-purpose it can be. Let’s review a list of MoClo ...Assembly of multiple Level 0 parts into TUs. Level 2: Assembly of multiple Level 1 TUs into multigene constructs...restriction enzyme and a ligase are used to assemble multiple DNA fragments together in a one-tube reaction....individual DNA parts, transcriptional units, and multigene constructs. Let’s start by examining individual...

...end of the multi-cloning site, and the reverse primer is located in the 5’ end of the multi-cloning site... one of the oldest of these techniques and is a multi-step process consisting of digestion, purification...

...Plasmids for multi-input CRISPR/Cas9 genetic circuits References: Nielsen AA, Voigt CA. Multi-input CRISPR... and Alex Nielsen describe what’s possible when multi-input CRISPR/Cas genetic circuits are linked to ...

...HRP-RANbodies are, however, compatible with most multi-labeling immunohistochemical protocols where either.... Chicken IgY domains are especially useful for multi-color staining of tissue because they have limited...primary/ labeled secondary antibody. Compatible with multi-labeling protocols when other antigens are detected...RANbodies compared to antibodies. Staining with multiple HRP-RANbodies is indistinguishable since they ...cultured cells and tissue sections Great for multicolor staining of tissue since it won’t cross-react...

...primer and plasmid design. Examples:  “multiplex design”, “multi-method design”, and “single-method design...indels at a single locus, usually the target site. Multi-locus asseEENdb database ssment would be required...between tools. Examples: “single-target design”, “multi-target design”, “off-target aware”, “high mismatch..., “empirical design”, “single-PAM design”, and “multi-PAM design”. Trends: By the end of 2014, tools began... common utility functions are batch design or multiplex features. However, tools aiding in primer design...

... coli, SLIC is a cheap, standardized, and rapid multi-part DNA assembly method. Read on to learn how to...Gibson assembly. SLIC is a standardized method for multi-fragment DNA assembly, and its low cost makes it... Figure 2: Schematic of multicomponent assembly with SLIC. Multiple fragments are PCR amplified to...assembly, assembling fragments in multiple steps to avoid using multiple fragments that share homology in...BioRender.com.   SLIC also opened up a new world of multicomponent assembly, as fragments with overlapping sequence...issue is sequence similarity. If fragments in a multicomponent assembly have 5′ or 3′ sequence homology to...harnessing the power of DNA repair, you can assemble multiple fragments without the need for specific restriction...

...vectors with various resistances. Comprehensive multi-cloning site compatible with standard cloning using...use the same restriction enzymes for cloning so multiple constructs can be made and tested at once. Return...

...luciferase in plants Diego Orzaez 118069 MLRV Multiple Multiple Multi-luciferase reporter vector including transcriptional...Nano-lanterns) to allow for BRET-based in vivo imaging of multiple cellular processes, biosensors, or optogenetic...

...Figure 3: Multiplexed insertions with Type I-F INTEGRATE using a single plasmid containing a multi-spacer...insertions and/or deletions, to multiplexed gene knockouts (Vo et al., 2020), to multi-copy genomic integration...by expressing multiple crRNAs in parallel (Fig. 3), multiplexed insertions into multiple distinct targets...in type I-F INTEGRATE. The requirement for multiple Cas proteins to form the Cascade targeting complex... report of VchINT, three separate plasmids and multiple promoters were used to express the necessary components...exhibit something called target immunity, whereby multiple proximal insertions with the same transposon exhibit...insertions of even very large genetic cargo, at one or multiple desired genomic loci. Many applications of these...

...in a single “one-pot” reaction to form complex, multi-insert modular assemblies that enable biosynthetic...conditions. The ability to construct more complex, multi-fragment assemblies, as shown in this work, will..., the BsaI site is no longer present, allowing multiple fragments to be assembled in an ordered, pre-determined...

...Spaghetti Monster probes at Addgene Uses of smFPs 1. Multi-channel connectomic tracers Fluorescent proteins...antibodies. GFP emits light in only a single channel. In multi-color imaging experiments, the second color is usually...antibodies, making them difficult to use in a multicolor/multichannel imaging set-up. In contrast, smFP tags ...can have low affinity for these tags and even multimeric tags are frequently insufficient for detection...the visible spectrum, researchers can also use multiple FPs in single experiments and visualize numerous.... smFPs are literally composed of FPs fused to multiple epitope tags. Indeed, to create the smFPs Looger...specific reactivities making them more useful for multicolor imaging experiments than standard FPs. The Looger...

...for multi-drug resistance: definitions vary in the literature, but generally organisms with multi-drug...happen to possess resistance genes survive and multiply. Factors contributing to the development of resistance...

...species and/or different IgG subclasses of 1° for multi-color staining Can be expensive Nanobodies   ...before application to the sample. Staining with this multi-color staining workflow yields similar localization...), which allows for the following to occur: A) Multiple polyclonal secondary IgGs can bind to different...sites per antibody. C) IgG’s are big enough that multiple labels (green circles in figure 2 below) can be...of a tag or fusion with a reporter enzyme. For multicolor staining, careful consideration must be used ...Antibodies Bivalent Polyclonal Can be labeled with multiple dyes Commercially available Not recombinant...sacrifice Good tissue penetration Can be labeled with multiple dyes Greater resolution due to lower label displacement...