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We narrowed to 399 results for: POR C

Showing: 201 - 220 of 399 results

  1. Tetracycline Inducible Expression

    Type
    Collection
    ...opens in a new window) Krueger, C., Pfleiderer, K., Hillen, W., & Berens, C. (2004). Tetracycline derivatives...vector to express Tet-On 3G transactivator under the c-Fos promoter Tet-On 3G rtTA Bong-Kiun Kaang 20342 ... N., Schambach, A., Galla, M., Maetzig, T., Baum, C., Loew, R., & Schiedlmeier, B. (2011). Retroviral ...elements like the H1 RNA promoter. And, most importantly, the TetR protein has been modified to provide...identify the amino acid residues of TetR that were important for tetracycline-dependent repression. Mutating...efficacy by fluorescence, use Plasmid #35626 as a reporter for cDNA target. rtTA3 Ptet, miniCMV Howard Gu...

  2. A Needle in a Base-Stack: Cas9 Structural Biology

    Type
    Blog Post
    ...similar nuclease folds found in other proteins. The C-terminal domain (CTD) of the protein also belongs ...https://doi.org/10.1038/s41586-022-04470-1 Cofsky, J. C., Soczek, K. M., Knott, G. J., Nogales, E., & Doudna...Sternberg, S. H., Redding, S., Jinek, M., Greene, E. C., & Doudna, J. A. (2014). DNA interrogation by the...it is limited to only 20 bases, or why it’s so important to choose a target sequence with a nearby protospacer-adjacent... lobes laying open like a book (Figure 2). The portion of the CTD that recognizes the PAM is so flexible..., but they improve cutting efficiency and are important for robust activity in cells (Jiang & Doudna, ... guide RNA stabilizing the DNA-binding protein portion of the CTD, ready to check for PAM sequencines....

  3. Performing In Vivo CRISPR Screens Using the FITS Approach

    Type
    Blog Post
    ...References and resources Milling, L. E., Markson, S. C., Tjokrosurjo, Q., Derosia, N. M., Streeter, I. S....screens have been used to interrogate T cells, an important immune cell type, in more complex and physiological...

  4. QC Sequencing Technologies at Addgene

    Type
    Blog Post
    ...reactions, where a fraction of one of the four bases (A, C, T, or G) is supplied as a dideoxyribonucleoside triphosphate...regions of interest, or complex sequences such as G-C rich areas, secondary structures, or repetitive regions... technology, but Oxford Nanopore is one of the most popular options. Nanopore sequencing deviates from...pulled through the pore via motor proteins. When each nucleotide passes through the nanopore, the electrical...3: Overview of Nanopore sequencing technology. Created with BioRender.com. Nanopore can provide longer...sequencing methods (Figure 3). It uses (aptly named) “nanopores” that are attached to the flow cell membrane and...-read sequencing. At Addgene, we routinely use Nanopore technology to sequence our packaged viral DNA ...

  5. Proximity Labeling: A Powerful Tool for Protein Complex Purification and Proteomic Mapping

    Type
    Blog Post
    ...these methods a try.  References:  Branon, Tess C., et al. "Efficient proximity labeling in living cells...1837. PubMed PMID: 30676735. Kim, D. I., Jensen, S. C., Noble, K. A., KC, B., Roux, K. H., Motamedchaboki...the mitochondrial calcium uniporter (Martell et al., 2012). It is important to note that the biotin-phenol... of exogenous biotin allowing for more precise temporal control.  Find plasmids for TurboID and miniturbo... where two proteins are fused to complementary reporter proteins that can only assemble when in close ... ascorbate peroxidase as a genetically encoded reporter for electron microscopy." Nature biotechnology...proteomics approach to monitor the composition of spatiotemporally defined protein complexes." Nature communications...

  6. Hot Plasmids Spring 2024

    Type
    Blog Post
    ...specificity of GFP expression for the indicated ssAAV5s. C) Quantifications showing high microglia transduction... microglia-targeting AAV plasmids here! Serrano, C., et al. (2023). Simple and Highly Specific Targeting...2024). They developed a new prime editor that incorporates La’s N-terminal domain, PE7 (Figure 2).  PE7...high microglia-specific gene expression when incorporated in an AAV vector (Figure 5). Efficiency and ...including in human retinal organoids and live mice. Importantly, REVeRT avoids the introduction of extraneous...associated with viral vector-based therapies. REVeRT supports advanced gene expression regulation, enhancing...

  7. CRISPR Plasmids - Prime Edit

    Type
    Collection
    ...gRNAs By Species Mammalian Bacteria Drosophila Plant C. elegans Yeast Zebrafish Xenopus CRISPR Resources ...which a reverse transcriptase (RT) is fused to the C terminus of Cas9 H840A nickase. The fusion enzyme ...the PBS of the pegRNA and the desired edit is incorporated into the DNA by reverse transcription. The edited...

  8. The Fluorescent Vegetables in Aptamer Soup

    Type
    Blog Post
    ...due to poor folding and thermal instability at 37°C (Ouellet et al., 2016) and requires a tRNA scaffold...imaging single molecules and for live-cell imaging in C. elegans. Other groups have used Mango to study RNA-protein...split aptamer is composed of a “Top” and “Bottom” portion, each fused to a separate piece of RNA. Fluorescence...photostable, and were presented in the study to support the uniqueness of the photostability of the Corn-DFHO...growth as RNA biology, we are looking forward to reports on the next additions to aptamer soup! References...

  9. Tips and Tricks for Using Golden Gate Modular Cloning (MoClo)

    Type
    Blog Post
    ...frozen glycerol stocks. It is very important that these stay frozen at -80 °C, since multiple rounds of freezing...; CDSns, coding sequence with no stop codon; TAG, C-terminal tag; TER, terminator. Created with BioRender.com... perfect Our first tip, and arguably the most important, is to take some time to sit down, plan your cloning...

  10. Getting the Most from Your Lentiviral Transduction

    Type
    Blog Post
    ...into single-use aliquots, and stored at -80°C. Some reports suggest rapidly freezing virus in a dry ice... for less than a day, lentivirus can be kept at 4°C. For long-term storage, viral preps should be divided...protein attaches to cell surface receptors, other important binding events take place during the course of...

  11. Plasmids 101: E. coli Strains for Protein Expression

    Type
    Blog Post
    ...conditions. Many recombinant proteins express better at 30°C or room-temperature, which is accomplished by growing...growing your culture to the desired density at 37°C and reducing the temperature or moving it to a bench-top...added to the growth medium. Induction timing is important, as you typically want to make sure your cells... optimal because of the high cell-density they support; however, minimal media supplemented with M9 salts...

  12. Tips for CRISPR Gene Editing in Mice

    Type
    Blog Post
    ...entire gene? Perhaps you are more interested in the c-terminus and would like to target that region. Or ... can be cloned directionally into the BbsI site. (c) The pCAG-EGxxFP target plasmid was co-transfected...expressing Cas9 and single guided RNA." Scientific reports 3 (2013). PubMed PMID: 24284873. PubMed Central...

  13. Delivery Methods for Generating iPSCs

    Type
    Blog Post
    ...Zhi Ying Chen, Robert C. Robbins, Mark A. Kay, Michael T. Longaker, and Joseph C. Wu. "A nonviral minicircle...a Sendai Virus-Based Vector Using a Nonstructural C Protein and Endogenous MicroRNAs." Plos One 11, no...of Different Methods for Generating iPSCs. One important note: in this table, reprogramming efficiencies...high is based on the reprogramming efficiencies reported in González et al. A percentage is not listed ... be used to package reprogramming factors. An important milestone of reprogramming is the silencing of...cells for the presence of transgenes is still an important control. Additionally, Sendai virus doesn’t rely...comprised of OCT4, SOX2, LIN28, NANOG, and a GFP reporter. Minicircles are removed from cells by dilution...

  14. Plan Your Experiment

    Type
    Collection
    ...Whitehead, E. H., Guimaraes, C., Panning, B., Ploegh, H. L., Bassik, M. C., Qi, L. S., Kampmann, M., &...Delivery Validate Genetic Edit References CRISPR ( C lustered R egularly I nterspaced S hort P alindromic...25408407 Dixit, A., Parnas, O., Li, B., Chen, J., Fulco, C. P., Jerby-Arnon, L., Marjanovic, N. D., Dionne, D...testing multiple pegRNA designs is particularly important for prime editing. CRISPR Intereference : Target...Predicted On-Target and Off-Target Activity Another important factor to keep in mind when designing your gRNA...In addition to off-target activity, it is also important to consider factors that maximize cleavage of ...generally perform by chemical transfection or electroporation. Expression can be transient, or you can generate...

  15. Targeting HIV-1 with CRISPR: Shock and Kill or Cut it Out?

    Type
    Blog Post
    ...’s most common diseases. References Zhang Y, Yin C, Zhang T, Li F, Yang W, Kaminski R, Fagan PR, Putatunda...Gendron P, Zhu W, Guo F, Cen S, Wainberg MA, Liang C. CRISPR/Cas9-Derived Mutations Both Inhibit HIV-1 ...infection, including both common infections and opportunistic infections not usually seen in healthy individuals... genome had been excised, leaving only a small portion of the LTRs joined together. Clonal Cas9/gRNA-expressing...populations were also immune to HIV-1 reinfection. Importantly, no off-target effects were detected. Kaminski...patients. Although this is an exciting finding, it’s important to note that these HIV-1 patients were ART-naive...Cas9. While these results are discouraging, it’s important to note that these experiments were done in cell...

  16. Evolution of Brainbow: Using Cre-lox for Multicolor Labeling of Neurons

    Type
    Blog Post
    ...Sato T, van Es JH, van den Born M, Kroon-Veenboer C, Barker N, Klein AM, van Rheenen J, Simons BD, Clevers...melanogaster. Hadjieconomou D, Rotkopf S, Alexandre C, Bell DM, Dickson BJ, Salecker I. Nature Methods. ...the 1980s, is one of the most important ways to spatially and temporally control gene expression, especially...ten colors (see table to the right); in various reports, Sanes and Lichtman have observed from 90-160 distinct...undergone recombination. This default state was disproportionately expressed by a majority of neurons, limiting... 8 colors. The use of AAV provides spatial and temporal control without the need for germline modification...refinements of the Brainbow technique will provide important insights into the complicated physical organization...

  17. CRISPR History and Development for Genome Engineering

    Type
    Collection
    ...and others) Drosophila Plants (monocots and dicots) C. elegans Yeast ( S. cerevisiae and S. pombe ) Zebrafish...Zebrafish Xenopus References Barrangou R, Fremaux C, Deveau H, Richards M, Boyaval P, Moineau S, Romero ...23287718 Dalvai M, Loehr J, Jacquet K, Huard CC, Roques C, Herst P, Côté J, Doyon Y. 2015. A Scalable Genome-Editing-Based...Adamson B, Villalta JE, Chen Y, Whitehead EH, Guimaraes C, Panning B, Ploegh HL, Bassik MC, Qi LS, Kampmann ...libraries can be used to identify genes that are important to a given phenotype. Current libraries are available...

  18. Fluorescent Proteins 101: History of Fluorescent Proteins

    Type
    Blog Post
    ...expressed GFP in exogenous organisms (E. coli, and later C. elegans) for the first time. After Chalfie’s work...responsible for encoding GFP, and, in 1992, he reported its sequence. Soon after that, in 1994, Prasher...he lived! His hunch paid off, and he was able to report new fluorophores like DsRed from Discosoma sp. ...provided by the studies in Tsien lab, covered a portion of the color spectrum that GFP and its derivatives...reach. Nobel Prize for fluorescent proteins The importance of fluorescent proteins was recognized in 2008...

  20. The Pleiades Promoter Project

    Type
    Collection
    ...., Chen, Y., Cheng, J. C., Chopra, V., Docking, T. R., Dreolini, L., D'Souza, C. A., Flynn, E. K., Glenn...intron-lacZ/NLS References Portales-Casamar, E., Swanson, D. J., Liu, L., de Leeuw, C. N., Banks, K. G., Ho...plasmid webpages. These plasmids are described in Portales-Casamar et al., 2010 . List of Pleiades MiniPromoters...MiniPromoters MiniPromoter Source Gene Construct Reporter Ple2 ADORA2A pEMS1142 EGFP/NLS Ple3 ADORA2A pEMS1143...
Showing: 201 - 220 of 399 results