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We narrowed to 971 results for: lars

Showing: 161 - 180 of 971 results

  1. What's New in CRISPR - November 2021

    Type
    Blog Post
    ...Endogenous eNcapsidation for cellular Delivery (SEND). They demonstrate SEND as a modular system for packaging...and specify a large intervening deletion. Not only can this method precisely program large genomic deletions.... pyogenes CRISPR/Cas9 system is that it is too large to be effectively packaged into AAV particles for...  with an active nuclease.  Read the paper in Molecular Cell Find the CasMINI plasmids Efficient CRISPR...plasmids   Selective endogenous encapsidation for cellular delivery of RNAs By Mike Lacy The retroviral-like...endogenous Gag homolog, cargo RNA, and fusogen in a modular system for delivering gene editing tools into cells...(epegRNAs) protect the RNA from degradation by cellular exonucleases and broadly improve editing efficiency...

  2. Split Fluorescent Proteins for Studying Protein-Protein Interactions

    Type
    Blog Post
    ...Christopher Grefen’s lab or explore the MoBiFC modular bimolecular fluorescence complementation toolkit from...available to assess protein-protein interactions. One popular approach is to fuse a protein of interest to each...Figure 1). This method is generally known as Bimolecular Fluorescence Complementation (BiFC). It can provide...proteins are split between strand 10 and 11 to yield large (1-10) and small (11) fragments. In several cases...Next, let's look at a round-up of some new and popular split-FP plasmids from our catalog to help you ...) for use with GFP(11) Multiplexed labeling of cellular proteins with split fluorescent protein tags. ...

  3. Early Career Researcher Toolbox: Finding Relevant Papers

    Type
    Blog Post
    ...jumping into the discussion! Semantic Scholar Semantic Scholar is an AI-backed literature search developed...Finding research papers is not particularly hard. There are millions of them. The real challenge is finding...also set up alerts for new publications from a particular author. Saving papers Papers can be saved to ...accounts can help you find papers related to a particular topic or that were published by the lab down ...follow journals related to your field of research or larger journals that publish about a broad range of topics...Institute for Artificial Intelligence. Semantic Scholar analyzes publications and figures using machine...relevant to your search terms. Key features Semantic Scholar has several ways to focus your search results. ...

  4. Viral Vectors 101: AAV Variables That Matter

    Type
    Blog Post
    ...different system (Issa et al., 2023). This can be particularly burdensome in the brain where many different...broad tropism (the ability of a virus to enter a particular cell type), but there are differences in tropism...using chemogenetic tools, where modification of cellular activity outside of your experiment site or circuit... offer the most flexibility for expression in a large number of cell types and are useful if you're going...several different systems or want to transduce a large number of different cells. Promoters such as chicken-beta-actin... Selective promoters, such as synapsin, glial fibrillary associated protein (GFAP), or tyrosine-hydroxylase...vectors in the central nervous system. Frontiers in Molecular Neuroscience, 7. https://doi.org/10.3389/fnmol...

  5. Troubleshooting and Optimizing a Western Blot

    Type
    Blog Post
    ...check its subcellular location and solubility. If it’s membrane-bound or located in a subcellular compartment...about which type of gradient can help answer your particular question before running one.  Good on gradients...strengths. If that’s not working, consider using a subcellular fractionation kit or a lysing protocol developed...enough to get good separation in your ladder, particularly in the sizes around your protein of interest...overheating remains an issue, which can happen with large proteins, try lowering the voltage and using a longer... leftover sample) to fill the relevant wells. Similarly, if you load a gel and then forget to start it...proteins going completely through the membrane or larger proteins not transferring completely onto the membrane...

  6. Plasmids 101: 5 factors to help you choose the right cloning method

    Type
    Blog Post
    ...options could be good choices if you have a particularly large number of inserts to assemble. You could ...figure out what system might work best for your particular purposes. We’re here to help with that! Four ...Cost Difficulty Efficiency For cloning in particular, another important consideration is: The number...post, we will focus on a high-level comparison of popular cloning methodologies, with a special emphasis ...cloning approach should you use if the budget is particularly tight? Most cloning processes will require a...and overall you’re looking at a price of a few dollars per reaction for traditional restriction cloning...obtained for about $70, resulting in a price of a few dollars per reaction. By comparison, Gibson assembly can...

  7. Plasmids 101: What is a plasmid?

    Type
    Blog Post
    ...the lab. Addgene’s Molecular Biology Reference includes information about molecular cloning, how to choose...updated Apr. 14, 2020. Any newcomer who joins a molecular biology lab will undoubtedly be asked to design..., or construct a plasmid. A plasmid is a small circular piece of DNA found in bacterial cells, and someone... will provide you with an overview of general molecular biology knowledge and techniques, and empower ... At their most basic level, plasmids are small circular pieces of DNA that replicate independently from...plasmids as they grow, scientists can easily make large amounts of plasmid to manipulate and use in later...versatility, safety, and cost-effectiveness enable molecular biologists to broadly utilize plasmids across ...

  8. Fluorescent Proteins 101: Fluorescent Biosensors

    Type
    Blog Post
    ...presence or changing cellular conditions. Other types of biosensors To focus on a particular gene or protein...be expressed. Are you looking at an intracellular or extracellular molecule? By fusing a biosensor to ... domain, biosensors can be targeted to a particular cellular compartment such as the nucleus, mitochondria...luminescent and fluorescent reporters being especially popular. We’ll give you an introduction to fluorescent ...by Olivier Pertz’s lab. Other biosensors track cellular characteristics like pH, redox state, and voltage...mitochondria, endoplasmic reticulum (ER), vacuole or cellular membranes. Examples include blue flame mitochondria-targeted...calcium sensor CMV-mito-GEM-GECO1  and blue flame extracellular glutamate sensor pCMV(MinDis).iGluSnFR. Picking...

  9. Plasmids 101: Secondary Nanobody Toolbox

    Type
    Blog Post
    ...strength. This signal strength is due in large part to their larger size (~10-15 nm), which allows for the...together without forming large oligomeric complexes. Antibodies are also too large to be expressed in bacteria..., than antibodies for all of the above listed molecular techniques, but they can also be expressed in ...antibody structures. Functional comparison Strong but Large: Advantages of Antibodies One of the main reasons...being big is a double-edged sword. An antibody’s large size creates more distance between the target antigen...ability to penetrate tissue. Because antibodies are large and bivalent, primary and secondary antibodies cannot... Poor tissue penetration Less resolution due to larger label displacement Require separate incubation ...

  10. Antibodies 101: Validation

    Type
    Blog Post
    ...researcher toolkit due to their ability to bind to molecular targets with high affinity and specificity. However... does not mean that is all it will bind to - particularly when you start using complex samples such as...Validation (Uhlen, et al., 2016). This group proposed 5 Pillars of antibody validation which represent five experimental...performs as expected. One of the benefits of these pillars is that they are intentionally generalizable and...summarizing the IWGAV five proposed antibody validation pillars. Created with BioRender.com.     Orthogonal...your antibody can distinguish between two very similar proteins. Additionally, they require variable expression...1) other strategies may not be feasible and (2) large-scale proteomic and transcriptomic datasets already...

  11. FLEx Technology and Optogenetics: Flipping the switch on gene expression with high spatial and temporal resolution

    Type
    Blog Post
    ...partner to optogenetics to understand the cellular and molecular mechanisms of the brain in vivo. Many...energy conversion or the induction of intra- or intercellular signaling pathways (Ernst et al., 2014). Microbial...adeno-associated viruses (rAAVs) have become increasingly popular for gene delivery in the central nervous system...ventral tegmental area (VTA) of the brain. In particular, they found that optogenetic stimulations of ...animal rhodopsins: structures, functions, and molecular mechanisms." Chemical reviews 114.1 (2013): 126...functional photoreceptor for phototaxis in the unicellular eukaryote Chlamydomonas." Nature 311.5988 (1984...Central PMCID: PMC4987841. Lodish, Harvey et al. Molecular Cell Biology. New York: W. H. Freeman, 2000. Bookshelf...

  12. Advancing Biology with Zebrafish: Genetic Tools for Developmental Studies and More

    Type
    Blog Post
    ... Zebrafish are also popular outside of developmental biology, and are particularly well-suited for neuroscience... development. Why have zebrafish become such a popular model organism? Zebrafish (Figure 1) were initially... the function of these genes in fish will help clarify their roles in humans. The sequenced genome also...Zebrafish mutants In the 1990’s, two revolutionary large-scale random mutagenesis screens produced ~1,500...TALENs, and more recently CRISPR/Cas9. Another popular targeted gene deactivation method in zebrafish ...exploits the bacteriophage integrase phiC31. The popular UAS/Gal4 and Cre/Lox gene regulatory systems have...tail. Compared to the opaque mouse embryo, where similar processes occur over days inside the mother, the...

  13. Twenty Years of Sharing: Expanding Accessibility

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    Blog Post
    ...our popular blog series even more accessible by collecting and formatting all 101s on a particular topic...execute a step can provide needed clarity (Table 1). Our most popular video, however, is “What is a Plasmid...offering temperature-sensitive materials, we noticed a large percentage of these shipments arrived in India at...help with sharing information through the use of popular Chinese social media platforms.  As you can imagine...expertise in a specific country can quickly reduce a large number of barriers. Though we haven’t solved every...about them. As early as 2007, we began grouping similar tools together in what would eventually become ... right for their experiment.     Table 1: Most popular guides, protocols, and collections of 2023. ...

  14. Avoiding the Dark Side of Fluorescent Protein Fusions with mOX FPs

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    Blog Post
    ... to learn that FPs are often poorly suited for cellular compartments other than the cytoplasm. Generally...eukaryotic) proteins localize to chemically distinct subcellular environments, including the organelles that compose...mitochondria, lysosomes or they are secreted into the extracellular milieu. Many of the resident proteins within...function of resident ER chaperones and may impact cellular function or possibly viability. FP misfolding .... The moxFPs overcome the inherent problems of popular fluorescent proteins and were re-engineered by ...residues and N-glycosylation consensus sequences in popular FPs. This generated a palette of inert green, cyan...most inert solutions for imaging in a variety of cellular compartments including the secretory pathway (...

  15. Special Delivery: Fluorophore Targeting for FRET Studies

    Type
    Blog Post
    ...how a protein of interest actually behaves. How large is it? What parts of it move when you feed it substrate...does it have and how do they come off and on in a cellular environment? If these are pressing issues in your... proteins, lipid bilayers, organelles or other cellular impediments. FRET has been used in many applications...LanthaScreen) (Degorce et al., 2009), measuring intracellular calcium (Miyawaki, Griesbeck, Heim, & Tsien,...excellent primer on FRET and how to measure it). Molecular mailbox: How to get your fluorophore delivered...(a single mailbox at a specific house in your molecular city). However, the acceptor doesn’t need to go.... If the acceptor fluorophore is getting to a molecular mailbox that is close to the donor, faulty delivery...

  16. Enabling Precision Functional Genomics with the Target Accelerator Plasmid Collection

    Type
    Blog Post
    ...we developed assays to test large numbers of cancer variants across a large number of genes as systematically...structure. Sometimes this provides a great deal of clarity. But, often these methods don’t agree. Confusing...characterization For many years, we’ve hoped that large scale laboratory-based functional experiments that...poorly classified function and a well established cellular signaling gene, thus providing a starting point... us from testing this hypothesis: There was no large scale, broadly available collection of mutant cDNA...reference cDNA clones mapping to well-annotated cellular pathways). It wasn’t clear whether gene-agnostic...gene-agnostic bioassays could accurately test for molecular signatures of variant function and cancer phenotypes...

  17. Viral Vectors 101: Integrase-Deficient Lentiviral Vectors

    Type
    Blog Post
    ...transgene is incorporated into a large complex of both viral and cellular proteins and trafficked to the...Lentiviral vectors have been a staple in molecular biology for over three decades. Widely used across...This integrated viral genome is then part of the cellular DNA… forever!  This can be an advantage for researchers...enzymes that are proteolytically processed from the larger protein: PR (protease), RT (reverse transcriptase....  The envelope plasmid typically encodes the vesicular stomatitis virus G protein (VSV-G), which facilitates...proviral DNA is poised for integration, two types of circular episomes can be generated through either homologous... your own IDLVs is nearly identical to that of regular lentiviral vectors. The only difference is that...

  18. CRISPR 101: Cytosine Transversion Editors

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    Blog Post
    ...the end if you ever need to double-check your vocabulary! A brief review of base editing tools First, ...an inhibitor of base excision repair (BER), the cellular process that catches mutated U’s in DNA and ideally...Researchers in the Xueli Zhang lab were making similar observations around the same time. (The papers ... journal on the same day.) This team created a similar construct that they called a Glycosylase Base Editor...the base editor. Black arrows indicate natural cellular processes. Created with BioRender.com. Beyond...abasic site. But how that site is filled depends on cellular factors. Multiple labs tried to identify beneficial...repairing a GBE-derived abasic site requires more cellular factors binding to DNA. However, ABEs were improved...

  19. Antibodies 101: Flow Compensation

    Type
    Blog Post
    ...antibodies may have similar emission spectra, meaning they will emit fluorescent light at similar wavelengths ...fluorescence for both at similar levels, meaning that your populations will ‘look’ similar on your flow plot....between the two of them because they emit at a similar wavelength. However, if your flow cytometer reads...is if the names of the fluorescent markers are similar, you will need to compensate between them (although...against each other, you should be able to draw perpendicular lines that intersect your populations. If you...population needs to be shifted, consider your perpendicular lines. In this case, the population on the left...needs to shift towards the X axis to become a perpendicular vertical line.     In general, most spillover...

  20. Fluorescent Tagging of Endogenous Genes with SapTrap

    Type
    Blog Post
    ...associated with cellular organelles, scientists have been able to study many cellular processes, including... for studying the organization and function of cellular systems. FPs have been used to track protein localization... localization, cell structure, intracellular trafficking, and protein turnover rates. Additionally, by...trafficking. Although FPs have enabled discovery of many cellular mechanisms, there are some limitations to working... ultimately misinterpretation of the protein’s cellular role. One way to avoid the pitfalls of overexpressed...SapTrap The Jorgensen Lab recently developed a modular plasmid assembly toolkit called SapTrap to improve... with C. elegans, several groups have designed modular toolkits to assist with tagging genomic loci in...
Showing: 161 - 180 of 971 results