... Depositor Collections MORF Feng Zhang - Multiplexed Overexpression of Regulatory Factors (MORF) Collection...Plasmids ID Plasmid Gene/Insert Pooled Library The Multiplexed Overexpression of Regulatory Factors (MORF) pooled...

...Which FP Should I Use? Blog: How to Choose a FP For Multi-Color Imaging Blog: Avoiding the Dark Side of Fluorescent...

...4-COMP-his-stop Type II expression CD69-His-bio Multi TM expression CD36-bio-His Plasmid collections exist...proteins are expressed as both monomeric baits and multimerised enzyme-tagged preys. Ectodomain inserts: The...

...Freemont Lab MoClo-YTK : Assembly of single and multi-gene constructs for S. cerevisiae - Dueber Lab Yeast...Golden GATEway cloning system - Wittbrodt Lab MultiSite Gateway Kit : Creating three-part fusion proteins... in genome editing of bacteria. CRISPathBrick Multiplex Plasmid : CRISPR expression in E. coli - Koffas...

...vectors containing several cassettes and generate multi-construct transgenic plants - Jan Lohmann. MoClo... seed selection and multiplex CRISPR/Cas9 assembly for fast generation of multiple Arabidopsis mutants... of useful plant CRISPR resources include: A Multipurpose Toolkit enables targeted, specific modification...CRISPR/Cas9 system for convenient, high-efficiency multiplex genome editing in monocot and dicot plants - Yao-Guang...expression vectors: A modular plasmid assembly kit for multigene expression, gene silencing, and silencing rescue...Infrastructure Kit for hierarchical assembly of plant multigene constructs - Johannes Stuttmann. MoChlo: Modular...

...following in 2015. Class 1 (Multi-subunit effector complex) Class 2 (Single multi-domain effector) Type I ...not only simple, but also scalable. Multiple gRNAs targeting multiple loci can be easily expressed in the...genome engineering, offering flexibility and easy multiplexing/scaling. Prokaryotes have long utilized CRISPR...the tool’s specificity, orthogonality, and multiplexibility in various species, as well as the development...being able to target every locus in every genome. Multiple Cas9 orthologs have also been characterized. Staphylococcus...distinct PAM sites that do not cross react, allowing multiple Cas9s to be used together. Researchers have engineered...Cpf1's small size also makes it suitable for multiplexing and use in AAV. Scientists have also developed...

...Figure 1: Schematic of multiplex immunofluorescence approaches. A) Sequential Multiplex IF, where one target...just want to make some really beautiful images? Multiplex IF may be just what you need! How does it work...work? There are several ways to achieve multiplex IF. One of the most common approaches is to use sequential...before labeling the next target. B) Simultaneous Multiplex IF, where all the primary antibodies are combined...   Figure 2: Examples of simultaneous multiplex IF labeling of rat brain samples from the James...your antibodies When selecting antibodies for multiplexed IF, many of the same considerations apply as ... to avoid secondary antibodies altogether and multiplex with direct IF, using primary antibodies that ...

... Such plasmids are known as multimers, and they are the result of multiple plasmid copies combining in...in a process known as plasmid multimerization. Plasmid Multimerization Early studies on plasmid topology...various multimeric forms (e.g. concatemers, dimers, trimers, tetramers). Since then, multimerization has ...plasmid multimers function just as well as monomers. However, due to their larger size, multimers may exhibit... detecting plasmid multimers. Digests themselves cannot be used to detect multimers, since restriction...frequency of multimerization, and in some cases completely prevent the formation of plasmid multimers (Bedbrook... pUC multimerization was reduced by growing seed stocks at 30°C compared to 42°C. Screen multiple colonies...

...use a multitude of techniques including co-transfection of two or more plasmids, the use of multiple or ...co-expression of multiple unique genes, you can start with a plasmid that has multiple cloning sites flanking...within the same multicistronic vector, effectively making a construct that expresses multiple unique genes...Co-expression of multiple genes is valuable in many experimental settings. To achieve this, scientists...promoters, or the creation of bicistronic or multicistronic vectors. Unlike promoters which will create...transcripts for each gene that is expressed, multicistronic vectors simultaneously express two or more ... so in this blog post we’ll cover basics of multicistronic vectors: why they are useful, how they work...

...Unless you use a system for processing a continuous multi-gRNA transcript, each gRNA must be expressed from...sites. Modifying multiple genes at once. Using multiplexed gRNAs can target multiple locations in the ...CRISPR makes it easy to target multiple loci - a concept called multiplexing. Since CRISPR is such a robust...you add multiple gRNAs on one plasmid. Sound good? Addgene has many tools to help you multiplex - we’ll...Drosophila, and zebrafish!   Why use multiplexed gRNAs? By expressing multiple gRNAs on the same plasmid, you...clone and transfect multiple promoter-gRNA constructs in order to target multiple sites! In this post,...we'll cover Cas9 multiplexing options, but also check out our blog post about multiplexing with Cpf1.   ...

...simultaneous multi-gene editing with a single crRNA array. Are you interested in using Cpf1 multiplexing in your...for multiplex genome editing.  For a few reasons, Cpf1 is a simplified system for editing multiple targets...protospacer Multiplexing CRISPR-Cas9 options before Cpf1 crRNA array Prior to this new Cpf1 multiplexing method...allow for expression of multiple gRNAs (i.e. Csy4 cleavage sequence, tRNAs, multiple individual promoters...Cpf1 counterparts. Table 2. Cas9 Multiplexing Options  Multiplexing Method Delivery Method Advantages...Xie et al  Multiplexing crRNA expression with Cpf1  The key advantage of multiplexing crRNA expression... chart below for a summary of multiplexing efficiency in vivo. Multiplex gene editing with CRISPR-Cpf1...

...scalability and efficiency of multiplex gene editing. Moreover, existing multiplex strategies have been facing...developed DAP arrays for efficient multiplex base editing (MBE) and multiplex prime editing (MPE) with only...Schematic of multiplex gene editing with DAP array   Use DAP arrays can enable efficient multiplex gene ...natural CRISPR locus of a bacteria host encodes multiple guide RNAs (gRNAs) on a single array to target...leveraged such host-defense mechanisms to enable multiplex gene editing in a variety of cells and organisms...from a single array, has been a top choice for multiplex gene knockout or regulations, it lacks proper ...contains a double-win for both efficiency and multiplexity, with these constraints? The Gao lab first ...

...identification. While the Zhang lab controlled their multiplicity of infection so that a single TF was delivered...to each cell, future applications could deliver multiple TFs to each cell, allowing for combinatorial overexpression...dysregulation event. Diseases are often complex, with multiple layers of dysregulation from either genetic mutation...

...Takeshi Imai from Kyushu University. Stochastic multicolor labeling is a powerful solution for discriminating...neuronal reconstruction. To achieve stochastic multicolor labeling, Brainbow used the Cre-loxP system to...fluorescent protein (XFP) genes in a transgene. When multiple copies of the transgene cassette are introduced...introducing a mixture of XFP genes. Stochastic multicolor labeling is possible when each of the XFP genes...Richi, Marcus N. Leiwe, and Takeshi Imai. "Bright multicolor labeling of neuronal circuits with fluorescent...

...Synthetically Phosphorylated Isomers (iSPI) Described in: A multi-purpose, regenerable, proteome-scale, human phosphoserine...orthogonal translation system to make singly or multiply phosphorylated proteins (Pirman et al., Nat Comm...

...et al. "Molecular basis for inhibition of AcrB multidrug efflux pump by novel and powerful pyranopyridine...

...biologists (Bohórquez lab), who together developed multifunctional fibers, embedded, and validated simultaneously...adaptable tool that could continuously monitor multiple systems. Ideally this tool would be wireless, ...optical stimulation of its associated fibers across multiple channels and can also record tissue temperature..., Chandrakasan, A., … Anikeeva, P. (2023). Multifunctional microelectronic fibers enable wireless modulation...

...will be necessary to improve its multiplexing capacity. Multiplexing is key due to the natural redundancy...knockout, or multiple gRNAs may be used to target genes with overlapping activity. Multiple gRNAs can also... Addgene Other Techniques for gRNA Multiplexing Multiplexed and programmable regulation of gene networks...;  to observe a phenotype, the modification of multiple genes is often necessary. Guide RNAs (gRNAs) are...editing efficiency. One previous strategy for multiplexing gRNAs used the RNA nuclease Csy4 from Pseudomonas... test the application of this system to gRNA multiplexing, Xie et al. constructed PTGs containing 8 gRNAs...mutations at all five loci. This highly efficient multiplexing further illustrates the potential of PTGs. Applications...

... step in choosing fluorescent proteins for your multi-color imaging experiment is to be aware of what ... multi-color imaging...cell imaging experiments is the ability to follow multiple fluorescently tagged species simultaneously. To...do so with fluorescent protein labels requires multiple fluorescent proteins whose excitation and emission...fluorescent proteins Commonly used filter sets for multicolor imaging include ones designed for CFP, YFP, and...compartments. I hope this sheds some light on multicolor imaging with fluorescent proteins. With the right...

...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...