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Showing: 361 - 370 of 587 results
  1. CRISPR 101: Validating Your Genome Edit

    Type
    Blog Post
    ...faster when using a dual guide system.   Fig. 2: Size screening for large deletion events.   ... Biol 17: https://doi.org/10.1186/s13059-016-1012-2 Find more CRISPR Resources at Addgene: Read our CRISPRGuide...number of cells have been edited. For many experiments, the end goal is to generate a clonal cell line, so...designed your gRNA and introduced it into your target cells with Cas9. Hooray! Now it’s time to make sure your...blog post we’ll explain how to verify that your cells were appropriately edited for your desired mutation...important to know a few things about your target cells and the desired edits. First, you will need to know.... Validating knockout mutations When a knockout cell line is desired, any mutation that introduces a ...
  2. Retrograde AAV viral preps

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    Collection
    ...Calcium sensor Ting 83899 pAAV-mDlx-GCaMP6f-Fishell-2 mDlx GCaMP6f expression under the control of the mDlx... of Cre, and expresses NLS-EGFP in Cre-positive cells. Control Harvey 137164 pAAV-nEF-Con/Fon/Von eYFP...
  3. Plasmids 101: Stringent Regulation of Replication

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    Blog Post
    ...the chromosome, ending replication.   Figure 2: Prokaryotic chromosomal replication. Adapted from...Microbiology and Molecular Biology Reviews: MMBR, 62(2), 434–464. https://doi.org/10.1128/MMBR.62.2.434-464.1998...replicating too quickly can slow cell growth or even kill the host cell. For stringently controlled plasmids...All plasmids rely on their host cell's replication machinery in order to replicate—but not always to ...be synchronized with the replication of the host cell's chromosomal DNA.  Plasmids are said to be under... stringent control when they depend on the host cell’s initiation proteins to start their own replication... occurs at the appropriate time—for example, the cell must have enough nutrients available to complete...
  4. Your Top Requested Plasmid in 2015!

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    Blog Post
    ... 24336571. PubMed Central PMCID: PMC4089965. 2. Sanjana, Neville E., Ophir Shalem, and Feng Zhang. "Improved...both Cas9 and your gRNA of interest to mammalian cells. It was designed to have increased lentiviral titer...knock-out thousands of genes in mixed population of cells as part of pooled library screens (1,2). The human...pooled libraries targeting both mouse and human cells are avaliable for distribution. To learn more about... includes work probing mTOR signaling in cancer cells, deciphering the mechanism of matrix metalloproteinase...of the many plasmids being distributed to and accelerating research in labs all around the world. We thank...Genome-scale CRISPR-Cas9 knockout screening in human cells." Science 343.6166 (2014): 84-87. PubMed PMID: 24336571...
  5. Empty Backbones - Choosing Your Perfect Plasmid Backbone

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    Collection
    ...backbone with MS2 loops at tetraloop and stemloop 2 and EF1a-zeo resistance marker pLenti CMV/TO GFP-Zeo...and display of proteins on the extracellular surface of S. cerevisiae cells His-tagged versions of pFastBac... the elements needed for replication in the host cell and are designed for a user to insert a gene of ..., or express the insert they carry in the target cell. These vectors allow you to test the function of....1-mCherry - Expresses mCherry tag in mammalian cells pBI-MCS-EGFP - Tet-inducible Find...or Fusion Protein Tags and fusion proteins are excellent tools for further understanding the function... as GFP, which allows you to visualize the cellular localization of your protein. Additionally...
  6. CRISPR 101: Which Cas9 Do I Choose for My CRISPR Experiment?

    Type
    Blog Post
    ...sequence is unique compared to the rest of the genome.  2) The target is present immediately upstream of a Protospacer...utilized allowing for PAM flexibility. Fig. 2: Prime editor engaging target DNA. The reverse transcriptase...Generating a no-fuss genetic knockout  Knock-out cells or animals are engineered by introducing Cas9-mediated...transcription by blocking its initiation. In mammalian cells, robust repression requires targeting dCas9 along...study essential genes whose knockout would cause cell lethality. The Krüppel associated box (KRAB) is ...repressing target genes in a variety of species and cell types can be found on Addgene’s website. Activating...activating target genes in a variety of species and cell types can be found on Addgene’s website. ...
  7. Podcast: Interview with Addgene's Founders

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    Blog Post
    ...Intro Chapter 1 - Who are Addgene's Founders? Chapter 2 - The process of founding Addgene, where the idea ...co-founder of Addgene. Melina received her Ph.D. in Cell Biology from Harvard University, where she studied...Plasmids Find Plasmids for FRET Find Plasmids for Subcellular Localization ...
  8. Inntags: Innovative Protein Epitope Tagging

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    Blog Post
    ...tagging." Nature methods (2015). PubMed PMID:26322837. 2. Stadler, Charlotte, et al. "Immunofluorescence and...that inntags IT5, IT6, and IT10 expressed well in cells and did not form undesirable aggregates. Using budding...tags. Impressively, IT6-GFP did not reduce yeast cell size or viability compared to untagged GFP overexpression...multiple other proteins. Again, little to no change in cell morphology was observed with IT5 and IT6 fusions...and HA tags had especially detrimental effects on cell size and growth rate, indicating that they compromised...correlation for protein localization in mammalian cells." Nature methods 10.4 (2013): 315-323. Pubmed PMID...
  9. Advice on Career Paths and the Green Card Process for International Researchers and Entrepreneurs

    Type
    Blog Post
    ... both substantial merit and national importance; (2) The foreign national must be well positioned to advance...outstanding achievements in your specific academic field, (2) that you have three 3 years of research experience..., Casseus Law suggests a mix of experts including 2 dependents –people who have worked with you i.e. your...This post was contributed by guest bloggers Lauren Celano of Propel Careers and Rachel Casseus, Esq. Founder...below. An international or national award for excellence in your field. Membership in a professional organization...counsel. Many thanks to our guest bloggers Lauren Celano of Propel Careers and Rachel Casseus, Esq. Founder...resident green cards and citizenship.   Lauren Celano, CEO Propel Careers. Lauren is passionate about...
  10. Countdown to Halloween @Addgene

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    Blog Post
    ...Halloween is only 2 days away and here at Addgene we couldn’t be more excited! As a small, tight-knit...teamwork, and competition. And our Halloween celebration this Friday will combine all of these. Addgenies...
Showing: 361 - 370 of 587 results