We narrowed to 20 results for: CRE-ER

...localize them to the secretory pathway, a dark pool that is mislocalized to the ER can be detected with...blocked from proceeding through the secretory pathway, and retained in the ER. FPs have been undeniably powerful... FP fusion with a resident endoplasmic reticulum (ER) protein instead localizes to the cytoplasm? Or more...absolute position requirements. For example, a KDEL ER retrieval sequence only functions at the extreme ...potential to interfere with the function of resident ER chaperones and may impact cellular function or possibly...complex-localized fusions from properly exiting the ER and fluorescing (see Figure 2) [5].  When using FPs...compose the secretory pathway, endocytic vesicles, mitochondria, lysosomes or they are secreted into the ...

... the ER lumen reaches the sub-millimolar range, much higher than in the cytosol. Thus, for the ER you ... expression, and compatible with constitutive or Cre-dependent expression. These sensors have also been...measuring the events-of-interest. Sensors suitable for ER measurements include: CEPIA (additional versions ... to simultaneously measure calcium changes in the ER, cytosol and mitochondria), the Cameleon-based FRET...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 ...

...fluorescent protein to the Endoplasmic Reticulum (ER). If homodimerization occurs (most likely in an antiparallel...used as positive and negative controls for proper ER structure. Based on these data, we conclude that ...tagging. On the other hand, mScarlet-I shows correct ER labeling, which is good evidence that mScarlet-I ...proteins are available, and their number is steadily increasing. Of course, you want the ‘latest and greatest...

... lab's SARS-CoV-2 sybodies! References Geertsma ER (2013) FX Cloning: A Versatile High-Throughput Cloning...Kuhn BT, Bräuer P, Newstead S, Dawson RJP, Geertsma ER, Seeger MA (2020) Generation of synthetic nanobodies...Hetemann L, Gera J, Gmür S, Spies P, Gygax D, Geertsma ER, Dawson RJ, Seeger MA (2018) Synthetic single domain...to, like ones that are hidden in the molecular crevices proteins on the cell surface. Figure 1: ...threonines at positions that can be randomized for creating the libraries (Zimmermann et al., 2018). Find ...protein (sybody), ribosome, and mRNA. During the screening process, displayed sybodies that bind to the biotinylated...the candidate sybodies in their coat protein to screen for binding to the target protein. The protocol...

...control without modification - either Cre is there or it’s not. Cre-mediated possibilities for site specific...control FLP and Cre expression have been developed. Read on to learn more about FLP-FRT, Cre-lox, and how...the control of Cre recombinase are used in combination with mouse lines that express Cre only in the cell...estrogen receptor (ER) ligand binding domain (LDB) to the C-terminus of FLP or Cre (pCAG-CreERT2 #14797). These...Tamoxifen inducible Cre versions can be used in these experiments. This elegant combination of Cre, FLP, and inducible...about Cre-Lox Check out Retrograde AAVs for Neurscience Research Resources on Addgene.org Find Cre-Lox.... Tamoxifen then binds to the ER-LDB domains of the fusions allowing Cre/FLP translocate to the nucleus...

...HRB (1987) A C-terminal signal prevents secretion of luminal ER proteins. Cell 48:899–907. https://doi....S, Pelham HRB (1986) An hsp70-like protein in the ER: Identity with the 78 kd glucose-regulated protein...

...the nucleus, mitochondria, endoplasmic reticulum (ER), vacuole or cellular membranes. Examples include... biosensor creation, and custom FRET biosensors can be created using the cpFRET kit created by Olivier...Robert Campbell’s lab display calcium-dependent increases in fluorescence of up to 11,000-fold. FRET biosensors...biosensors. Blue flame “cameleon” calcium sensors created by Roger Tsien’s lab contain two fluorescent proteins...calcium binds, it triggers CaM/M13 interaction that increases FRET transfer between the fluorescent proteins...

... Figure 2: Two-color super-resolution imaging of ER and mitochondria reveals dynamic organelle interactions...interactions. a) Labeling scheme. b) COS-7 cell ER labeled with Halo-Sec61β with SiR-CA (magenta) and outer...no wonder that self-labeling protein tags are increasingly popular. Embrace the flexibility and switch ...

...Structure Plasmids: ER mCherry-Sec61 beta BFP-Sec61 beta BFP-KDEL Rtn4a-GFP (tubular ER) Microtubules ... their synthesis in the ER to their arrival in their target compartment (e.g. the plasma membrane, the...stably localized in the donor compartment (e.g. the ER) and a reporter protein of interest (Figure A). Franck...and interactions between the endoplasmic reticulum (ER), mitochondria, cytoskeleton, and endosomes. These...upon these findings by screening for additional activation domains that increase the expression of target...Synchronization of secretory protein traffic in mammalian cells The journey of secretory proteins, from their... mutagenized and screened for speed and voltage sensitivity. This hierarchical screen led to the generation...

.... In tamoxifen-inducible Cre (Cre-ER) systems, Cre is fused to the ligand binding domain of estrogen receptor...the Esr1 domain that blocks Cre expression. Then, Cre is expressed and Cre-mediated recombination of floxed...that regulates Cre expression. The most common drug used to induce the expression of Cre is tamoxifen. ...doxycycline mediated activation of Cre to either induce or repress Cre expression using TetON or TetOFF ...using Cre-lox Read our genome engineering blog posts Resources on Addgene.org Find more about Cre-lox ... vector, creating the floxed gene segment. Crossing of mice containing a floxed gene to a Cre-expressing...mice, conditional knockout/knockin mice, use the Cre-lox recombination system to express the transgene...

...nucleus, nearly all cysteines are reduced; in the ER and Golgi apparatus and outside the cell, most cysteines...assembly tools, alongside criteria for Click Biology. Created with BioRender.com.   Principles of Click Biology...Protein (MFP) through rapid covalent coupling. Created with BioRender.com.   Table 1: A handful of...system, adds Tetracycline (Tet) transactivator to increase fluorescent protein expression C234-E25: attB2r-SNAP-attB3...Cas9 plasmids with N- or C-terminal PCV2 HUH-tag; increase homology-directed repair efficiency by linking...linking the repair template VirD2 HUH-tag For creating DNA-protein conjugates rHUH HUH-tag (RNA) Covalent...

... PHO8 is synthesized at the ER, delivered to the vacuole via the secretory pathway, and then cleaved to...In both cases, red fluorescence in the vacuole increased with longer exposure to autophagy-inducing conditions...

...postsynaptic AMPA receptor and is endocytosed, the Cre recombinase payload activates a floxed reporter gene...synapses. In addition, ATLAS itself could be expressed Cre-dependently to allow tracing exclusively from genetically... components (nucleus, endosomes, lysosome, golgi, ER, inhibitory synapses, and cytoskeleton components... 5: Mechanism of ATLASCre. VAMP2 = synaptobrevin 2; At = ALFA-tag; BACE = β-secretase; AF = AMPA.FingR...TnpB) or a gRNA (Cas9). Drawings not to scale. Created with BioRender.com. Steven Jacobsen and Jennifer...potent retrograde labeling of projection neurons. Created with BioRender.com. Addgene has tested these...Huang, et al. (2025) under CC BY-NC-ND license, created with BioRender.com. When delivered by AAV ...

...A-induced perturbation in biological processes in ER-negative HEK293 cells." PLoS One, 2014. 9(6): p. ...making their surrounding media acidic through the secretion of lactic acid (a by-product of cellular metabolism...

...to label proteins moving to and from the nucleus, ER, mitochondria, stress granules, and more. For added...enable the creation of CAR-T cells directly within the body. Overall, DIRECTED can increase on-target ...reprogram delivery vehicles like lentiviral vectors, CreVLPs, and eVLPs for in vivo and in vitro applications.... The Ting lab recently made two deposits that increase the utility of this technique.  By engineering... O’Neill The Cohen lab and colleagues recently created a system for recording the transcriptional history...resembling a “ticker tape” recording. The authors created externally-timed bands of color with HaloTag-iPAK4...thin sections of tissue, the Wu Lab is working to create a three-dimensional atlas of protein expression...

...References 1. Snapp, E.L., et al., Formation of stacked ER cisternae by low affinity protein interactions. J... second FP, often decreasing the fluorescent signal from the first FP and increasing the signal from the...an FP to a transmembrane protein significantly increases the effective concentration of the FP. The FP ...to a plane and generally only rotates in 360°, increasing the probability that two FPs will collide with...polymer formation as fusion protein concentration increases. The polymerized fusion proteins that form as ... His interests include the quality control of secretory proteins in the Endoplasmic Reticulum and live...

...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...how you can make CRISPR mouse model generation easi-er! Generating mice before CRISPR The process of generating...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-...

...R., & Okiyoneda, T. (2023). UBE3C Facilitates the ER-Associated and Peripheral Degradation of Misfolded...of protein activity. Together, these limitations create challenges for measurements needed to move discoveries...means to quickly answer research questions before creating knock-ins.  Figure 1: Schematic of HiBiT...Functional assays further showed that imported ADAR1 increased oncogenic editing of its most common target, AZIN1...UBE3C. Degradation of CFTR was measured through decreases in HiBiT signal, which was slowed by knockdown...factors present unique challenges for study. These secreted signaling proteins regulate vital cell and tissue...physiologically and endogenously relevant models, tools that increase confidence become more important. For many difficult-to-study...

...results, so you only need to pick your favorite (...er, whichever one uses assumptions that match your data...Welcome to our deep dive series, which aims to increase your understanding and technical proficiency with...temperature drops, single intensity should smoothly increase as the DNA amplicon transitions from single-stranded...finally, drop once more during a final temperature increase. If there is a single amplicon, there should only...c) A double peak indicating 2 amplicons. Image credits (respectively): Zuzanna K. Filutowska; Selma, 2019...can fit in a quick game of Connect Four! Image created in BioRender.   Passive Reference Dyes Whatever...

...this screen the authors were able to identify 7 previously unsuspected genes with known roles in ER-associated...loxP-stop-loxP cassette into the Rosa26 locus, allowing for Cre-dependent expression of a GMAP inserted gene. The...of a good screening experiment can be found on our blog post entitled “Genome-wide Screening Using CRISPR...Receptor-ome Expression and Screening via Transcriptional Output-TANGO (PRESTO-TANGO) to screen the NCC-1 library...and later create stable cell lines expressing it. The Church lab fixed this issue by creating a new dCas9...  CRISPR pooled libraries enable researchers to screen the entire genome for genes that regulate a wide...CRISPR knockout library and used this library to screen for genes involved in West Nile Virus (WNV) induced...