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We narrowed to 791 results for: Cre

Showing: 51 - 100 of 791 results

  1. Top Requested Lentivirus and AAV of 2016

    Type
    Blog Post
    Published
    Jan. 6, 2017, 3:56 p.m.
    ...Expression of hM4D(Gi)-mCherry from this virus is cre-dependent, meaning that once injected, the DREADD...DREADD will only be expressed in cells that express cre recombinase. Typically, this feature is used to attain...cell-specific expression in transgenic animals, where cre expression is restricted to a particular cell type...the first time, the Roth Lab recommends verifying cre expression prior to use. The lab additionally recommends...expressing cell lines, which can then be used both for screens (using pooled CRISPR knockout libraries) and individual...Secondly, you can use a LentiCas9-Blast viral prep to create a single stable Cas9-expressing cell line and then...monoclonal Cas9-expression line prior to performing a screen or even a single gene knockout. Top AAV: pAAV-hSyn-DIO-hM4D...

  2. Rabies and Neuronal Tracing

    Type
    Blog Post
    Published
    May 29, 2018, 1:51 p.m.
    ... For example, a researcher can deliver Cre-dependent TVA to Cre-expressing neurons in a transgenic mouse...Addgene Blog AAVs CREATed for Gene Delivery to the CNS and PNS Advanced Uses of Cre-lox and Flp-FRT - ...viral protein EnvA on its surface will only infect Cre-positive, TVA-expressing cells (Osakada et al., 2013...

  3. Viral Production at Addgene

    Type
    Blog Post
    Published
    Oct. 24, 2023, 1:15 p.m.
    ... the absence of Cre. mCherry expression alone was detected. pAAV-Ef1a-mCherry-IRES-Cre was a gift from...were transduced with either pAAV-Ef1a-mCherry-IRES-Cre (55634-AAVrg) alone at 1.7E6 viral genomes (vg)/cell...) alone at 1.1E6 vg/mL, or both. Two days later, Cre-dependent GFP expression was detected with direct...

  4. Plasmids 101: Inducible Promoters

    Type
    Blog Post
    Published
    Jan. 18, 2018, 2:34 p.m.
    ...Addgene depositors have developed heat shock-inducible Cre and Cas9 for easy genome engineering in species like... Plasmids 101: The Promoter Region Plasmids 101: Cre-lox Light-Switchable Two-Component Systems for E.... Tetracycline (Tet) Inducible Systems at Addgene Cre-lox Guide and Plasmids     ...cell growth media with glucose decreases cAMP and represses pBad, decreasing promoter leakiness.   Other ...

  5. Newly Updated AAV Data Hub!

    Type
    Blog Post
    Published
    Feb. 7, 2023, 2:15 p.m.
    ...interneurons can be labelled with an AAV1 Cre virus combined with a Cre-dependent AAV9 tdTomato virus in the...learn more about the AAV Data Hub, and hear the incredible AAV talks given by Connie Cepko and Ian Wickersham...

  6. Hot Plasmids - May 2022

    Type
    Blog Post
    Published
    May 26, 2022, 1:15 p.m.
    ...labeling Increased neuronal expression using Cre-mediated AAV Targeted Evolution method (M-CREATE) dCas9...-4.   Increased neuronal expression using Cre-mediated AAV Targeted Evolution method (M-CREATE) by: Brian...multiplexed version of the Cre-mediated AAV Targeted Evolution method (M-CREATE), in which one applies both...variants that display increased neuronal expression in the brain while having decreased expression in the ...zebrafish as a model organism, the Parton Lab recently created a flexible system that makes use of pre-existing...Image from Goertsen D, et al., 2022.  Find M-CREATE plasmids here!  Goertsen D, et al. Nature Neuroscience...

  7. Plasmids 101: The protein expression toolbox

    Type
    Blog Post
    Published
    Feb. 16, 2023, 7:05 p.m.
    ...control when you turn off expression for good, the Cre-Lox system offers inducible yet permanent gene excision...can be excised by intentional introduction of the Cre recombinase. This is a time-tested system but does...some permanent knock out plasmids? Check out our Cre vector and Murine LoxP vector.   Turn it off for ...!  Turn it on Turn it on forever Permanently increasing or turning on expression of a new protein is ... gRNA, to your gene of interest to temporarily increase expression. There are several flavors of activators...

  8. What's New in CRISPR - Spring 2019

    Type
    Blog Post
    Published
    March 21, 2019, 12:47 p.m.
    ...doxycycline, but it can also be regulated spatially with Cre-recombination. Read the paper in Frontiers in Molecular...Averof lab. It’s an alternative to the conventional Cre or Flp recombinases used to excise markers. It has...Cas12a inhibitors Using a bioinformatics and a screening approach, the Jennifer Doudna lab identified three...

  9. A Primer on Optogenetics: Introduction and Opsin Delivery

    Type
    Blog Post
    Published
    Sept. 10, 2015, 2:30 p.m.
    ...open-reading frame (DIO). Upon coexpression with cre recombinase (either from a mouse line or viral co-injection...type specificity dependent on the promoter of the cre recombinase while still taking advantage of the robust...an appropriate mouse line expressing a particular cre, viral expression of your opsin is no longer required...tracing with fluorescent proteins in nontransgenic and cre driver mice. Current protocols in neuroscience / ...: PMC4242517.  7. Madisen L, et al. A toolbox of Cre-dependent optogenetic transgenic mice for light-induced... us to show causality in neural circuits with incredible temporal and spatial precision (I’m not going...optogenetics has already been proven. Therefore, a concrete, well defined, and robust behavioral output is...

  10. Using AAV for Neuronal Tracing

    Type
    Blog Post
    Published
    Aug. 9, 2018, 1:04 p.m.
    ...often made Cre-dependent. If Cre-dependent vectors are used in transgenic animals that express Cre in only...labelling the subset of infected cells that also express Cre. Another advantage of AAV vectors is that there are... traced neurons with an AAV9 vector expressing a Cre-dependent fluorescent marker. Injection into a conditional...conditional mutant mouse brain that expressed Cre recombinase only in pyramidal cells of the hippocampal...PMID: 4107258. Lo, L., and Anderson, D.J. (2011). A Cre-dependent, anterograde transsynaptic viral tracer...

  11. 22 Hot Plasmid Technologies from 2014

    Type
    Blog Post
    Published
    Jan. 6, 2015, 5:21 p.m.
    ... short hairpin) and CRUSH (Conditional RUSH) use Cre-mediated recombination to turn on or off the expression...a dual-color RNAi “sensor” mouse strain in which Cre expression causes the induction of both DsRed fluorescence...Mice carrying the R26DsRedR; CRUSH-GFP; and Nestin-Cre alleles showed efficient GFP knockdown in eyes and... recombination events at the genetic level. Upon Cre/loxP recombination, each transgene expresses one ...Flp recombinase/FRT system in place of Brainbow’s Cre/loxP, allowing for simultaneous use of the two systems...SUMO tags in the FP sequence for separation from Cre-based Brainbow-expressing cells. Autobow plasmids... all-in-one versions which express self-excising Cre recombinase from the same transgene as the Brainbow...

  12. Hot Plasmids - August 2020

    Type
    Blog Post
    Published
    Aug. 18, 2020, 1:15 p.m.
    ...sensors include syn-jYCaMP1s (slow variant), the Cre-dependent syn-FLEX-jYCaMP1s, and the axon-targeted...syn-FLEX-axon-jYCaMP1s. Find these AAVs at Addgene Cre-dependent EYFP AAV in the new serotype PHP.V1 that...Find this AAV at Addgene New serotypes for the Cre-dependent Flpo recombinase pEF1a-DIO-FLPo-WPRE-hGHpA...biotechnology tool to optimize another. The authors have created a yeast-based directed evolution approach with ...your studies.  Find the plasmids to set up your screening systems at Addgene! Schematic of the evolution...following a DNA miniprep protocol!  First, the authors created the pMAK backbone by modifying pMAL-T-Avi-His/BirA...exported to the periplasm of bacteria where it can be secreted into the supernatant. Four plasmids containing...

  13. Addgene's Top 10 Blog Posts of 2015

    Type
    Blog Post
    Published
    Dec. 7, 2015, 6 p.m.
    ...in CRISPR genome editing.   2. Plasmids 101: Cre-lox Cre recombinase can be used to generate inversions...community members. In addition we’ve experienced incredible growth with our readership expanding from ~10,000...next revolution in biological research. We’re incredibly happy to see that one of our career posts has...protein of interest. 4. Plasmids 101: Blue-White Screening Getting a lot of empty vector contaminants in ...Addgenie Jessica Welch explains how Blue-White screening could be a potential solution. 5. Cpf1: A New ...

  14. An “elegans” Approach to Better CRISPR/Cas9 Editing Efficiency

    Type
    Blog Post
    Published
    Dec. 17, 2020, 1:30 p.m.
    ... the expression of a Cre recombinase contained within the knock-in. Including Cre-mediated marker excision...kilobases of additional sequence being introduced. Cre-mediated excision of the selective cassette minimizes...experimental manipulation. In some cases, injection of Cre is required, however the development of the self-...variable amounts of experimental manipulation and screening.  A co-selection strategy for CRISPR editing in... with inefficient sgRNAs in my initial direct screening efforts, which were similar to the approach detailed...low efficiency (0.13%) and laborious handling (screening 768 F1 animals). Recovering rare editing events...conditional-lethal mutation to identify edits with minimal screening effort. Selecting for repair of a lethal mutation...

  15. New and Upcoming Viral Vectors - June 2019

    Type
    Blog Post
    Published
    June 18, 2019, 1:15 p.m.
    ...recombinase that can be used in combination with Cre for intersectional control of specific neuronal populations... We now offer a Flp-expressing AAV, as well as a Cre-dependent Flp-expressing AAV. See our recombinase...

  16. New Neuroscience Tool: The iGluSnFR3 Glutamate Sensor

    Type
    Blog Post
    Published
    April 7, 2022, 1:15 p.m.
    ...CAG-FLEX: CAG promoter, FLEXed for expression in Cre-expressing cells hSynap: Human Synapsin-1 promoter...hSynap-FLEX: Synapsin-1 promoter, FLEXed for expression in Cre-expressing cells. hSynap-FLP: Synapsin-1 promoter...they created three affinity variants – A184S (increased affinity for glutamate), S72A (decreased affinity...as: iGluSnFR3 v82: increased affinity and slower kinetics iGluSnFR3 v857: decreased affinity and faster... forms of neuronal signaling and regulation. To create iGluSnFR, the Looger lab wedged GFP between two...to glutamate, conformational changes lead to an increase in fluorescence intensity from the inserted GFP...iGluSnFR, the Looger lab swapped eGFP for sfGFP to create SF-iGluSnFR. This improved variant has higher expression...

  17. PiggyBac-ing Through the Genome Editing Field

    Type
    Blog Post
    Published
    May 31, 2016, 3:30 p.m.
    ...piggyBac™, you could use recombinase technology (Cre/loxP) to remove the donor cassette. While this method...mutants [7], it is generally not as efficient as Cre recombinase. However, the use of negative selection...likely due to DNA toxicity. More importantly, unlike Cre recombinase, the PBx is highly specific, and has ...applications, including transgenesis, gene-trap screens, and gene editing. How does the piggyBac™ system...of the major uses of the piggyBac™ system is to create stable transgenic cell lines. The traditional method...site-specific nuclease (such as CRISPR/Cas9) to create a double-stranded DNA break. After DNA cleavage... editing? There are primarily two other ways to create specific point mutations by gene editing. First...

  18. Viral Vectors 101: An Introduction to AAV

    Type
    Blog Post
    Published
    May 30, 2023, 1:15 p.m.
    ... be achieved using specialized promoters, Flp or Cre dependent gene switches, and serotype selection. ...cell-specific gene expression and viral infection. Cre or Flp dependent switches control gene expression...

  19. Hot Plasmids - October 2020

    Type
    Blog Post
    Published
    Oct. 13, 2020, 1:15 p.m.
    ...targeting inhibitory or excitatory synapses or that are Cre-inducible, researchers can explore synaptic integration...preps at Addgene hSyn1-SIO-stGtACR1-FusionRed is a Cre-dependent, soma-targeting inhibitory channelrhodopsin...brighter in worms than split-sfCherry3. It was created by Maria Ingaramo, Jerome Goudeau, Cynthia Kenyon...Addgene The Cas Hybrid for Multiplexed Editing and screening Applications (CHyMErA) Knockout Libraries express...

  20. Simple CRISPR-based Epigenetic Editing: dCas9-directed DNA Demethylation

    Type
    Blog Post
    Published
    Dec. 14, 2021, 2:15 p.m.
    ... floxed dCas9 so that it may later be excised by Cre recombinase. Addgene has several great options: pLV...T2A-GFP for removable dCas9 expression and pLM-CMV-R-Cre. Selection for cells stably and highly expressing...are dioxygenases that oxidize 5-methylcytosine to create rare DNA bases that are recognized by distinct ...

  21. Fluorescent Proteins 101: Monitoring Cell Mobility Using Fluorescent Proteins

    Type
    Blog Post
    Published
    Aug. 15, 2017, 1:24 p.m.
    ...useful tool for marking and tracking cells is the the Cre/lox recombination system. A researcher can flank ...expression is turned on or off in cells expressing Cre. The result of a system like this is a mosaic of ...respectively. The permanent labelling of cells using cre-lox is advantageous when compared to photoactivable... In the past 15 years efforts have been made to create transgenic mice expressing fluorescent protein ...wavelength as it has been shown that even a small incremental change of the excitation wavelength can significantly...called Brainbow, the Ubow mouse strain has been created to fate map Langerhans cells and follicular dendritic...microscopy and new constructs have facilitated the creation of mouse lines that can be used in fate mapping...

  22. Multiplexed Capture of Promoter-enhancer 3D Chromatin Structures Using CRISPR

    Type
    Blog Post
    Published
    July 2, 2020, 12:40 p.m.
    ...More About Designing gRNAs Against Your Favorite CREs Here Additional Resources on Addgene.org Find ... cre... to make out the details. To increase the resolution, the Xu lab created a dCas9-based CAPTURE (CRISPR... smaller pieces. Sticky ends created are then ligated together to create loops of DNA. This is key to ...that, when bound by transcription factors, can increase a gene’s transcription. Sometimes enhancers are...method The original CAPTURE method requires the creation of a stable cell line that co-expresses three ...DNA into smaller pieces, and proximity ligation creates chimeric circles of DNA that are distant linearly...

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

    Type
    Blog Post
    Published
    Aug. 13, 2019, 12:58 p.m.
    ...bacteriophage integrase phiC31. The popular UAS/Gal4 and Cre/Lox gene regulatory systems have also been successfully...zebrafish transgenesis with Tol2 and application for Cre/lox recombination experiments." Methods in cell biology...two revolutionary large-scale random mutagenesis screens produced ~1,500 zebrafish mutants that are still...deactivation tools for zebrafish Random mutagenesis screens provided a major toehold into understanding how...et al., 2019). Generating zebrafish transgenics Creating transgenic organisms by introducing foreign DNA...one-eyed pinhead mutant from the random mutagenesis screens. Microscopy-based lineage tracing can help illuminate...

  25. Rett Syndrome: A History of Research and Therapeutic Outlooks

    Type
    Blog Post
    Published
    Nov. 19, 2020, 2:15 p.m.
    ...(Guy et al., 2007). They accomplished this using Cre-Lox mouse models in which the endogenous MECP2 gene...excision of the Stop cassette using tamoxifen-inducible Cre not only halted the neurological damage but completely...two MECP2 labs (Gail Mandel and Stuart Cobb) to create the Gene Therapy Consortium. Leveraging their combined...

  26. Hot Plasmids Spring 2024

    Type
    Blog Post
    Published
    May 23, 2024, 1:15 p.m.
    ... Ace2N-mNeon-A122D and Voltron2 are available as Cre-dependent AAV1 vectors and are suitable for extended...found that administering these AAVs with a dilute Cre-expressing vector resulted in a sparsely-labeled ...CRISPR screens By Emily Bentley Building on their CROPseq method for optical pooled CRISPR screening, Paul...Multiplexed perturbation and decoding in pooled CRISPR screens Improved prime editor PE7 Antibodies to facilitate...multiplexing approach to Cas9-based pooled CRISPR screens called CROPseq-multi (Walton et al. 2024). This...enrichment, single-cell sequencing, and optical pooled screens in human cells. Addgene offers vectors for CROPseq-multi...multiplexed perturbation and decoding in pooled CRISPR screens. bioRxiv 2024.03.17.585235; doi: https://doi.org...

  27. Delivery Methods for Generating iPSCs

    Type
    Blog Post
    Published
    April 17, 2018, 1:37 p.m.
    ...formation in iPSC-derived and iPSC-transplanted mice. Cre-deletable or Tet-inducible lentiviruses address some...considered safe for clinical applications. Excisable Cre-lox lentiviral vectors: This approach is the same...standard lentiviral vector, but transient expression of Cre recombinase allows for deletion of the inserted transgenes...insertional mutagenesis. Random integration also creates heterogeneous iPSC cell lines, which can complicate...insertional mutagenesis and random integration creates variation between iPSC clones. Retroviruses are...which is removed by culturing cells at 38℃, but screening cells for the presence of transgenes is still ...

  28. Viral Vectors 101: Systemic Capsids

    Type
    Blog Post
    Published
    Oct. 12, 2023, 1:15 p.m.
    ...Systemic capsids packaging Cre-dependent transgenes can also be paired with Cre-expressing transgenic mouse...with appropriate cell type-specific promoters or Cre lines. Spatial transcriptomics suggests that AAV-...Huber, N., Pasca, S. P., & Gradinaru, V. (2016). Cre-dependent selection yields AAV variants for widespread...Deverman, B. E., & Gradinaru, V. (2020). Multiplexed Cre-dependent selection yields systemic AAVs for targeting...have developed techniques like CREATE (Deverman et al., 2016), M-CREATE (Ravindra Kumar et al., 2020),...primates (NHPs). These engineered capsids have increased transduction efficiency in the CNS, with greater...

  29. Easi-CRISPR: Generating Knock-In and Conditional Mouse Models

    Type
    Blog Post
    Published
    April 5, 2018, 12:42 p.m.
    ...But CRISPRing typical mouse models, including creating Cre-dependent conditional alleles, has remained ...fluorescent proteins (EGFP, mCherry etc.), recombinases (Cre, Flp etc.), and tetracycline drug induction systems...plasmid pmScarlet_C1! After creating the T7-DNA template, you'd use ivTRT to create the ssDNA construct as ...CRISPR genome editing has made it easier to create knockout alleles in a variety of species, including...repair template is used. The original design for creating a conditional allele, or floxed mouse, from Yang... and inject the ES cells into a mouse embryo to create chimeric mice. This laborious process takes ~8-...product as a template for reverse transcription to create your ssDNA. Note that the T7 promoter will not ...

  30. Three Key Considerations for Precise Neuronal Targeting Using AAV Technologies

    Type
    Blog Post
    Published
    Nov. 27, 2019, 2:03 p.m.
    ...Combinations of classical genetic tools, for example Cre/loxP or Flp/FRT, with AAV vectors are common approaches... strategies Combining different approaches can increase the specificity of cell targeting even further...delivery of AAV at the injection site results in increased specificity of the targeted cell population.  ...are also developing ways to improve delivery by increasing transgene cargo capacity, enhancing anterograde...

  31. X-CHIME: Context Dependent Germline Knockout in Immune Cells

    Type
    Blog Post
    Published
    Jan. 16, 2024, 6:13 p.m.
    ... pXPR_070 Violet-excited GFP (vex) Lineage-Cre; Rosa26-Cas9 S-CHIME Knocks out pairs of genes...of genes in immune cells typically involves the creation of germline knockout mice, which is time-consuming...cells can be implanted in irradiated recipients to create mice with gene knockouts in their immune systems...2019). A CRISPR-Cas9 delivery system for in vivo screening of genes in the immune system. Nature Communications...

  32. Important Considerations When Using AAVs

    Type
    Blog Post
    Published
    June 13, 2017, 2:30 p.m.
    ...specific system for controlling gene expression such as Cre-lox, may be sufficient to overcome the transport ...ability to produce neuron specific transfection using Cre-lox, this new method could allow for labeling of ...AAV7, AAV8, AAV9 Lung AAV4, AAV5, AAV6, AAV9 Pancreas AAV8 Photoreceptor Cells AAV2, AAV5, AAV8 ...looks like at 3 days vs 7 days vs 14 days etc. To decrease the time until expression, it is possible to use...

  33. Genetically-encoded Sparse Cell Labeling - A SPARC of Innovation

    Type
    Blog Post
    Published
    May 21, 2020, 1:15 p.m.
    ...limiting amount of AAV are injected into the brain of Cre-expressing mice to trigger recombination and expression...site upstream of the stop cassette results in decreased efficiency of recombination and preferentially...

  34. Hot Plasmids and Viral Preps - July 2021

    Type
    Blog Post
    Published
    July 27, 2021, 1:15 p.m.
    ...in AAV8! Expand your imaging experiments with the Cre-OFF/FLP-ON vector 137128-AAV8: Ef1a-Coff/Fon-sRGECO...of biosensors could be used in high throughput screening for agrochemical compounds that have the potential...

  35. Hot Plasmids - October 2022

    Type
    Blog Post
    Published
    Oct. 6, 2022, 1:15 p.m.
    ...Two of their constructs are currently available as Cre-dependent, EF1ɑ-driven viral vectors (AAV1) - and...called JEDI-2P. Using a custom, high-throughput screening platform to apply saturating mutagenesis on 21... 1: Summary of the approach and outcomes of the screen for improved genetically-encoded voltage indicators... in-vivo imaging of voltage fluctuations. Image credit: Liu Z, et al. Cell. 2022.  Find JEDI-2p viral... Anti-c-Myc [9E10] page for more details. Image credit: Addgene.   Find anti-c-Myc antibody here! ...

  38. Tetbow: Bright Multicolor Labeling for Neuronal Tracing

    Type
    Blog Post
    Published
    Jan. 24, 2019, 2:24 p.m.
    ...stochastic multicolor labeling, Brainbow used the Cre-loxP system to express one of the three fluorescent...Addgene #104110 - #104112) (Figure 2). We’ve also created constructs with containing chemical tags such as...

  39. Even more elegant: Single injection CRISPR/Cas9 in C. elegans

    Type
    Blog Post
    Published
    July 7, 2015, 3:36 p.m.
    ...added flanking loxP sites and a heat-shock inducible Cre to generate a self-excising cassette (SEC). To make...have used PCR-based screening to detect mutants, but this method limits screening to a few hundred worms... less than eight hours worth of hands-on time. Creating a modular, selection-based gene tagging approach...rare events difficult. Dickinson et al. sought to create a new and efficient selection-based CRISPR/Cas9...construct will serve as the repair template plasmid to create fluorescent and 3xFLAG-tag fusion proteins. The...knock-ins. These candidate animals can then be further screened based on their Roller phenotype. If SEC is inserted...separates the targeted gene from its promoter, creating a loss-of-function allele as well as a promoter-FP...

  40. Quick Guide to Working with Drosophila Part 2: Controlling Gene Expression in Flies with Gal4/UAS

    Type
    Blog Post
    Published
    July 21, 2017, 12:48 p.m.
    ...modular expression systems exist (eg. LexA/LexAop and Cre/Lox systems) in case you want to express one gene...Drosophila geneticists use the Gal4/UAS system. This incredibly useful, yet simple system allows you to systematically...simpler, this modular system saves you from having to create a new fly line every time you want to express YFG...

  41. A Novice's Perspective on Neuroscience - SfN2017

    Type
    Blog Post
    Published
    Dec. 15, 2017, 2:07 p.m.
    ... Resources on the Addgene Blog Advanced Uses of Cre-lox and Flp-FRT - A Neuroscientist's View Interview...artificial intelligence (including a team that created a novel form of AI that can learn new tasks and...

  43. What's New in CRISPR - November 2021

    Type
    Blog Post
    Published
    Nov. 18, 2021, 2:15 p.m.
    ...modular system for packaging, secretion, and delivery of specific RNAs, such as Cre mRNA or Cas9 mRNA and sgRNA...for its small size. They were able to increase activity and decrease the gRNA size by intensively remodelling... engineered guide RNAs Yong-Sam Kim’s lab also created a miniature yet active Cas system starting with...

  44. Announcing the Winners of the 2021 Michael Davidson and Roger Tsien Commemorative Conference Awards

    Type
    Blog Post
    Published
    Aug. 5, 2021, 12:15 p.m.
    ...dual virus and the experimental approach. (B) CR-Cre mice demonstrate pain response due to blue light ...microcircuits. “In the longer term, this screening platform may help us to create high-performance sensors to enable...mechanical allodynia, a condition where a light touch creates a painful sensation. The dorsal horn of the spinal...virus expression in CR neurons of the spinal cord. Created with BioRender.com. Image from Haoyi Qiu.   ...indicators. Tian developed a video-based high-throughput screen for directed evolution of biosensors based on their... to engineer many types of biosensors.   This screening method allowed Tian to optimize a red‑shifted ... the improved voltage indicator coming from the screen to understand mouse cortical and hippocampal neural...

  45. Getting Started with Behavioral Neuroscience: Standardizing Experiments and Using Viral Vectors

    Type
    Blog Post
    Published
    Aug. 20, 2019, 12:31 p.m.
    ...expression levels of certain genes. A popular method uses Cre recombinase on transgenic flox animals to introduce...light or noise should be avoided as this will also increase stress levels and affect behavior. A fact to consider... also to minimize the damage of tissue due to increased pressure of the added liquid. The volume should...editing/deletion using CRISPR. In some cases, a decrease of the expression efficiency can be observed after...

  46. BeHeard Award 2018: Diseases of Glycosylation, Arginine Mutagenesis, & Neural Development

    Type
    Blog Post
    Published
    June 26, 2018, 1:33 p.m.
    ...delete the functional portion of the Nfix gene using Cre recombinase and to generate Nfix-negative neural ...burden of immune symptoms on CDG patients and to increase their quality of life by reducing their risks ...ligand binding sites in T4 lysozyme by deficiency-creating substitutions." Journal of molecular biology 277.2...

  47. Viral Vectors 101: Calcium Sensors

    Type
    Blog Post
    Published
    Aug. 10, 2023, 1 p.m.
    ... expression, and compatible with constitutive or Cre-dependent expression. These sensors have also been...binding sites per sensor (to decrease unnecessary calcium buffering and to increase sensitivity), and is a ...fundamental to muscle and nerve function, hormone secretion, blood clotting, and as we all know, a structural...illumination with violet light and (2) exposure to increased intracellular calcium. It has been applied to ...

  48. Lighting Up Cell Signaling with Photoswitchable Kinases

    Type
    Blog Post
    Published
    July 25, 2017, 1:20 p.m.
    ...Fluorescent Proteins Control Protein Expression with Cre-lox Control Protein Activity with GFP Resources ...possible to create other photoswitchable kinases? The general design of psMEK1 also worked for creating three.... The Problem:  There is no common method for creating optically controlled kinases. The Solution: In...et al, the Lin lab present a generic design for creating photoswitchable kinases and use MEK1 as a model...often hard to decipher because it’s difficult to create short pulses of kinase activity. Zhou et al studied...feedback on endogenous MEK1/2 phosphorylation by creating a short pulse of psRaf1 activity. They did this...just 5 minutes post-Raf1 inactivation, and then decreases. Treatment of the cells with a PP1/PP2A inhibitor...

  49. Quickest Way to Deposit Plasmids: The Deposit Spreadsheet

    Type
    Blog Post
    Published
    March 8, 2017, 3:15 p.m.
    ..., lentiviral, retroviral, AAV, RNAi, luciferase, cre/lox, yeast expression, worm expression, insect expression...and Organization where your plasmids were first created. From there, we’ll use the information in the ...

  50. 15 Hot Plasmids from 2017

    Type
    Blog Post
    Published
    Jan. 9, 2018, 3:02 p.m.
    ...light-dependent Gal4 transcriptional activation and Cre recombination. Find PhoCl plasmids at Addgene. Zhang... for easy removal of the resistance cassette via Cre recombination. Each knockout includes unique barcodes...natural ancestors), and screened through libraries of new RFP variants for increased fluorescence, brightness...polycistronic cassettes and discovered reprogramming discrepancies, which they traced to a 9 amino acid N-terminal... did not lead to the observed reprogramming discrepancies. The polycistronic piggyBac transposon vectors... protein PhoCl expands optogenetics toolbox Increasingly popular Optogenetics tools use light to alter...PMID: 28241009 CRISPR coselection with ouabain increases editing efficiency Blue flame depositor Yannick...
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