...epigenetic modifications across the genome) has been implicated in disease states, and targeting the epigenome...to specific DNA sequences. Non-editing CRISPR applications direct catalytically dead dCas9 fused to a variety...small DNA methyltransferase from the prokaryote Mollicutes spiroplasma (M. Sss1) (termed MQ1.) The Q147L...doi.org/10.1038/nbt.3199 Find plasmids from this publication at Addgene. Klann TS, Black JB, Chellappan M...doi.org/10.1038/nbt.3853 Find plasmids from this publication at Addgene.  Lei Y, Zhang X, Su J, Jeong M, ...10.1038/ncomms16026 Find plasmids from this publication at Addgene. Liu XS, Wu H, Ji X, Stelzer Y, Wu.../j.cell.2016.08.056 Find plasmids from this publication at Addgene. Maeder ML, Angstman JF, Richardson...

... tools for familiar applications. But how does one invent a tool for applications that don’t even exist... Andrew York, Maria Ingaramo, and their team at Calico Life Sciences recently set out to do just that ...phenomenon, improve the tool, and develop new applications for magnetoresponsive proteins. Motivated by...organisms) are opaque, which makes most biomedical applications of these tools difficult or impossible. But ...tool with magnets would enable new research and applications that don’t yet exist, but it wasn’t clear that...reproduced from Frank Hayward et al. 2024 under CC-BY license. They did more experiments to see if any small...reproduced from Frank Hayward et al. 2024 under CC-BY license.   Video 1: Fluorescence movie of E. coli ...

...Few Complicated Non-natural amino acids 1 Various Many Complicated   Applications Organic...labeled through click chemistry. Although these multi-component systems can be complicated to implement,...having to clone a whole new construct for every application. Plus, the photophysical properties of chemical...advantages over fluorescent proteins, opening up new applications.   Figure 1: Live-cell labeling strategies...adapted from (Bottanelli et al., 2016) under CC-BY license. Combining orthogonal self-labeling tags on...self-labeling tag and a fluorescent protein) enables applications like multi-color imaging (Figure 2). If you’...reused from (Ulrichs et al., 2023) under CC-BY license.   But these tags aren’t limited to fluorescent...

...intended application - Remember, just because your antibody performs as expected in one application does ...strategies for antibody validation in different applications. Here, I want to highlight the general suggestions...intentionally generalizable and not specific to certain applications or antibody targets. That said, not every strategy...will be suitable for every antibody or every application. The goal is to show that your antibody has been...strategy to use will depend on your intended application and available resources.     Figure 1...confident in your data. This is a good option for applications like immunohistochemistry on human tissues, ...alternative translation start sites or alternative splicing that could lead to some form of the target protein...

...suppliers, and public health and medical organizations are working to help educate the public about proper...containing a phage origin of replication that can be packaged into replication-incompetent phage particles...Beisel et al., licensed under a Creative Commons Attribution 4.0 International License.   Improving...essential processes: cell wall synthesis, DNA replication, or protein synthesis. Because they target such...mammalian viral vectors in that the vectors are replication-deficient. Thus, a very large number of phagemids...CRISPR antimicrobials combines them with both replication-competent phages and antibiotics. Phage therapy...bacterial host range. To further potential clinical applications, Ando et al. recently engineered synthetic phages...

...for both transient and stable gene expression applications in plants. This smaller, easily customizable...Agrobacterium are termed binary vectors because they must replicate in two species: Agrobacterium and E. coli. Like.... coli. However, they also need to be able to replicate in Agrobacterium to ensure that they are maintained... Thus, binary vectors contain two origins of replication, one each for E. coli and Agrobacterium, along...bases each. By switching the E. coli origin of replication and codon-optimizing the Agrobacterium ORI, they...constitutively active M24 promoter has been modified with duplicated enhancer domains and characterized in studies...these properties of pSiM24 are suitable for many applications, the plasmid can also be customized due to its...

...list of individual screens curated for the publication. Clicking on the screen name takes users to a display...favorite gene was described in a CRISPR screen publication. You see this mentioned in the results section...manually curated and annotated from more than 45 publications, beginning with the seminal research articles... makes it possible for researchers to query all public CRISPR results for any gene of interest. In addition...screen curated in ORCS is linked to the original publication via its PubMed identifier, which can also be ...screens of interest.   Figure 3: Each publication featured in BioGRID ORCS has a dedicated summary... supplemental data tables available with the publication are also provided for the user. This approach...

...for your application. Image reused form Doman and Pandey et al., 2023 under CC-BY 4.0 License. Find... CreVLPs, and eVLPs for in vivo and in vitro applications.   Figure 2: Modular features of DIRECTED...Strebinger et al., 2023, reused under CC-BY 4.0 License. Importantly, even commercial antibodies can...approach for experimental therapies and medical applications.  Find DIRECTED plasmids here! Strebinger, D...interactions or map a local proteome, but some applications have been challenging or inaccessible. The Ting...alkyne-phenol, to label proteins with an alkyne group for click chemistry reactions instead of biotin — a valuable...from Lin et al., 2023 bioRxiv, under CC-BY-NC 4.0 license. This system provides a unique new way to record...

...emission wavelengths, which are pivotal in their application in biotechnology (Grimm & Lavis, 2022). These... substrates/peptides and antibodies. Example applications of this include coupling to an antibody, whereby...typically bind to. Stability of fluorescence: For applications that involve ranges of pH, and significant lengths...light, is another factor to consider. If your application requires longer periods of exposure, you’ll want...have shown that different fluorescent ligands can elicit varying degrees of β-arrestin-2 recruitment to ...descriptions, or technical manuals. Specific applications and matching dye/ligand choices For effective...beyond the diffraction limit of light. Dyes like silicon-rhodamine or Janelia Fluor (JF) dyes have specific...

...to disparate physiological effects and complicate any metabolic study. The Ralser lab has created a unique.... Seeliger MA, Young M, Henderso MN, Pellicena P, King DS, Falick AM, and Kuriyan J. Protein Sci. 2005...brief articles highlight the main features and applications of a partiular plasmid or set of plasmids. We...Wu Lab library can be found in the associated publication. Ma, et al. Cell Rep. 2015. PubMed PMID: 26190106...and ubiquitous reporter assay. In their recent publication, Kroeze et al. describe their enhanced TANGO .... Each gene was chosen to represent the primary splice variant in 38 types of human cancer. Many of the...Sanger sequencing, respectively. The speed and simplicity of GMAP allows scientists to easily design, generate...

...Depending on the research question and downstream application, one or several of those techniques can be used...your flow analysis. However, certain downstream applications, like an in vivo adoptive transfer of those ...available fix/perm kits allow for labeling of cytosolic proteins, as well as proteins of the secretory...intranuclear proteins of interest, you can label cytosolic proteins alongside the intranuclear ones. You ...You do not need to use a separate fix/perm cytosolic procedure in that case.     Figure 3: For intracellular...or free amines of proteins. Depending on the application, they can be membrane-permeant or membrane-impermeant...are positive or negative for the dye. The main applications of dye stains in flow cytometry are cell proliferation...

... ITRs serve as the origin of replication and are required for replication, packaging, and vector persistence...viral rep (replication) and cap (capsid) proteins, which bind to the ITRs and initiate replication. ITRs are...binding site for Rep78 and Rep68 to initiate replication and a terminal resolution site that serves as... Rep proteins are important for viral genome replication, these mutations can affect transduction efficiency... of research is important for downstream AAV applications.  Tips & FAQs Due to their high GC content and... and hairpin structures, ITRs are prone to replication errors and disruptive mutations and truncations...mutations?  During propagation, mutants rapidly replicate in a growth culture and result in a high percentage...

...what we do not understand, and the complicated and unique metabolic processes of plants are being investigated...involved in plant synthetic biology. Challenge #1: Public perception of synthetic biology One of the biggest...difficulties that plant biotechnologists face is public perception of genetic modification (GM) and synthetic...benefits and future potential of the technology to the public in an open, responsible manner. At Revolution Bio...obstacles to plant synthetic biology In addition to public resistance, an overall limited understanding of...knockouts to better understand developmental, metabolic, and photosynthetic processes. Often the tools... intellectual property concerns – patents and licensing agreements. Developing a product without the use...

...constitutive splicing studies and has been optimized for high efficiency splicing. If intron splicing does not...potential applications of CRISPR/Cas9 in genome-engineering. One particularly attractive application involves...in order to avoid complicating effects from global toxicity due to loss of splicing. To this end, destabilizing... system widely accessible for many potential applications. Polstein LR & Gersbach CA, Nat Chem Biol ...amenable to both high and low throughput screening applications. The system can be readily modified to use different...cloning platform for imaging and flow cytometry applications Designing customized fluorescent reporters can...luciferase reporter system for detection of pre-mRNA splicing   The Gideon Dreyfuss lab has developed rapid-response...

...between bacteria can also have serious implications for public health. Bacteriocin plasmids: microbial...us! A plasmid is a small piece of DNA that can replicate independently from its host’s chromosome. Although...Some bacteria possess plasmid-borne toxins like colicins and subtilisins to kill other bacteria in the ...plasmids carrying genes for bacterial toxins like colicin typically also carry the necessary resistance genes...survive their own secretions. Pro tip! Historically, colicin-producing plasmids have been referred to as “Col...inducible expression systems that allow tuning of metabolic pathways. Fertility (F) plasmids: microbial sharing... common plasmid features, such as origins of replication and promoters, check out our other Plasmids 101...

...host genome, the virus is replication incompetent, meaning it can’t replicate and continue the infection... tools, thought was given on how to keep the replication of virus and its infectivity in check. Engineering...envelope plasmid), and the polymerase (Pol) for replication and Gag structural protein on yet a third plasmid... this safety feature prevents the virus from replicating and spreading in non-target cells, including ....    Viral cargo-specific safety   After replication competency, the second concern with virus safety... be disposed of following your lab’s biohazard policies. Administering Virus Many virus experiments involve... accident-prone researchers. Note that safety policies will vary slightly from organization to organization...

...most transposable elements (TEs) results in the duplication of this target site sequence at the site of insertion...This is similar to how retroviruses, like HIV, replicate. Class I TEs do not encode a transposase enzyme...enzyme.  Class I TEs are considered replicative since every time they jump they make a copy of themselves...retroviruses both in their structure and mechanism of replication. They contain two genes: gag and pol. The pol...they move. Most Class II transposons have a non-replicative “cut and paste” mechanism of transposition: they... it integrates, which produces a target site duplication (TSD). Created in BioRender.com.   Autonomous...transposition activity  Tol2 transposon plasmids    Applications of transposons Now that you’ve learned about...

...many CRISPR applications, and they’ve also had success in clinical gene therapy applications. Read on to...to learn more about the current (and future) applications of lentiviral vectors! Lentiviral vector history...original system, the possibility of creating a replication-competent virus via recombination between the...of recombination events necessary to create a replication-competent virus. Second, Tat is eliminated through...barcoding libraries have been used for complex applications like lineage tracing in heterogeneous tumors...developed integrase-deficient lentiviral vectors for applications that require the large genetic capacity of a...McGarrity GJ, Hoyah G, Winemiller A, Andre K, Stein D, Blick G, Greenberg RN, Kinder C, Zolopa A, Binder-Scholl...

...technique published every week! One of the newest applications is a colorful system that uses fluorescently...activity, catalytically dead Cas9 (dCas9) has many applications, the most well-known being transcriptional activation...characterizing the 4D nucleome. Future modifications and applications Ma et al. developed this technique using standard... et al. anticipate that the technique will be applicable to studies of cell cycle progression, epigenetics...to external stimuli. It could also have major applications to cancer, for example, the visualization of...Feb 23. Pubmed. Find the plasmids from this publication at Addgene. Dynamic imaging of genomic loci....2013.12.001. Pubmed. Find the plasmids from this publication at Addgene. A protein-tagging system for signal...

...Description Origin of Replication (ORI) DNA sequence which allows initiation of replication within a plasmid...plasmids are small circular pieces of DNA that replicate independently from the host's chromosomal DNA....virulence. All natural plasmids contain an origin of replication (which controls the host range and copy number...Minimally, lab-created plasmids have an origin of replication, selection marker, and cloning site. The ease... plasmids and the ability of plasmids to self-replicate within a cell make them attractive tools for the...plasmid by recruiting replication machinery proteins Antibiotic Resistance Gene Allows for selection of plasmid-containing...broadly utilize plasmids across a wide range of applications. Some common plasmid types include cloning plasmids...