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We narrowed to 856 results for: ELL

Showing: 541 - 560 of 856 results

  1. xCas9: Engineering a CRISPR Variant with PAM Flexibility

    Type
    Blog Post
    Published
    March 28, 2018, 6:52 p.m.
    ... in mammalian cells Hu et al. compared xCas9 activity with wt SpCas9 in mammalian cells, finding xCas9...ABE7.10, as well as improvements at NG and GAT PAMs. xCas9 3.7 Activity in Mammalian Cells Construct... percent editing efficiencies (edit frequency in cell population measured with NGS) are shown as percentages...

  2. Academic vs. Industry Postdocs

    Type
    Blog Post
    Published
    Nov. 13, 2018, 1:22 p.m.
    ...doing a brief academic postdoc. Why did I switch? Well, my long-term goal was to enter industry since I...” that you’ve sold out. You’ve given up your intellectual freedom and integrity. (Not the case, as I’ve...s ability to manage others and work as a team as well as their scientific abilities. At AstraZeneca, scientists...who get postdocs and do not support or manage them well are simply no longer allowed to have postdocs. In...Scientist at AstraZeneca. She has a background in cell and developmental biology and is applying this towards...

  3. Quick Guide to Working with Drosophila Part 1: Getting Started with Flies

    Type
    Blog Post
    Published
    July 13, 2017, 2:30 p.m.
    ...overexpress YFG in a Drosophila cell line. Knock downs in Drosophila cell lines are extremely effective....days after setting up the genetic cross). You can tell when pupae are ready to hatch the next day because...person. Animals are diploid except for their germ cells (sets of genomic material are contributed by mother...various fly reagents on the Bloomington website as well. Additionally, they have useful information on topics...

  4. PITChing MMEJ as an Alternative Route for Gene Editing

    Type
    Blog Post
    Published
    Feb. 23, 2016, 3:30 p.m.
    ... for Mapping Multiprotein Complexes in Human Cells.” Cell Rep. 13: 621-633. PMID: 26456817. Find plasmids...Chromosomes). Using their PITCh plasmids, GFP knock-in cell lines can be created in about a month and a half...locus-specific gRNAs Step 4: Select for puromycin resistant cells Step 5: PCR amplify and sequence the locus to verify...crispr.mit.edu. They tested their system in HEK293 cells, integrating the GFP-Puro cassette into the FBL ...

  5. Optogenetics + CRISPR, Using Light to Control Genome Editing

    Type
    Blog Post
    Published
    Sept. 3, 2020, 12:15 p.m.
    ...encoded tools, such as microbial opsins, to control cellular activities using light. In 2015, scientists combined...activator domain (VP64 or p65). Upon expression in the cell, the dCas9-CIB1 fusion binds to the target DNA sequence...activator domain VP64. Using this system in HEK293T cells to induce expression of human IL1RN, the researchers...protein with three red fluorescent tags in U2OS cells. Upon light activation, dCas9 is released within... silencing of transferrin receptor CD71 in HeLa cells.   Comparison of photoactivatable CRISPR strategies...Spatiotemporal Control of CRISPR/Cas9 Function in Cells and Zebrafish using Light‐Activated Guide RNA. Angew...

  6. CRISPR 101: RNA Editing with Cas13

    Type
    Blog Post
    Published
    July 31, 2020, 12:30 p.m.
    ...machinery, and could thus be used in non-dividing cells. Cas13 enzymes also don’t require a PAM sequence... RNA knockdown. In initial tests, Cas13b from Prevotella sep. P5-125 (PspCas13b) yielded 62.9% average... a maximum of 28% editing efficiency in HEK293T cells, as assessed using RNA-seq. Since the REPAIRv1 machinery...including reversibility and use in non-dividing cells. With the expanded targeting ability using both ...Functionally Orthogonal Type VI-A CRISPR-Cas Enzymes." Mol Cell. 66(3) (2017):373-83. PubMed PMID: 28475872. Find...

  7. Plasmids 101: Secondary Nanobody Toolbox

    Type
    Blog Post
    Published
    Feb. 27, 2018, 2:04 p.m.
    ...Nanobodies are like tiny antibodies which work just as well, if not better, than antibodies for all of the above...structure and function compare to IgG antibodies, as well as how to produce them for use in your lab. Structural...triple colocalization is possible even when staining cells with three IgG1 antibodies. Antibodies were individually...workflow yields similar localization patterns as cells stained for one target (compare nanobodies staining...–rabbit IgG secondary nanobodies. The Journal of Cell Biology. PubMed PMID: 29263082 Find plasmids from...

  8. Plasmids 101: Protein tags

    Type
    Blog Post
    Published
    Dec. 11, 2014, 4:26 p.m.
    ...not generally in a cell’s best interest to overexpress a protein. Energy and cellular resources are being...being spent to make something the cell doesn’t need to make. Eukaryotes and some bacteria deploy proteosomes...proteosomes to degrade what the cell might consider junk protein. Though there are a number of chemical and...same manner as it would have in the context of a cell. FLAG tags can be cleaved by enterokinase, which...

  9. Negotiating Work and Life: How to Find the Joy

    Type
    Blog Post
    Published
    March 20, 2014, 3:04 p.m.
    ...day care and school if you tell them they will enjoy it. If you cry and tell them they will miss you, of...having anyway) for enjoying your career and life as well as your family and friends. If there is any guilt... way more fun. And despite those who may wish to tell you so – towel corners do not need to match up exactly...example, say no to some of those birthday parties. Tell your mom you can’t make it to dinner this week. ...

  10. AAV Titers: Where Do They Come From And What Do They Mean?

    Type
    Blog Post
    Published
    Nov. 15, 2017, 3 p.m.
    ...particles that can transduce cells. Infectious titers are typically quantified by cell transduction assays. Wild-type... on three important factors. What can AAV titers tell you? Physical titer: the concentration of viral...arrowhead). Scale bar = 100nm. Image courtesy of David Bell and Svetla Stoilova-McPhie, Center for Nanoscale...Briggs HA, Gallichotte EN, Ruf IK, Hindson BJ, Vessella RL, Tewari M. Absolute quantification by droplet...

  11. Career Insights: Technical Support Specialist

    Type
    Blog Post
    Published
    Nov. 9, 2017, 2 p.m.
    ...Specifically, roles in technical support make excellent use of the skills developed by life science PhDs...issue was with a custom-made reagent to manipulate cells (manufactured by one of the larger biotech companies...patient enough to find the real issue, you will do well in a technical support role. What general characteristics...go wrong in a lengthy protocol. And customers may tell all sorts of details about the application, just...about novel developments in the fields of gene and cellular therapies.     Additional Resources on the Addgene...

  12. Designing Your Chalk Talk for the Academic Job Interview

    Type
    Blog Post
    Published
    March 12, 2019, 1:37 p.m.
    ...how do proteins X and Y regulate the Z pathway?" A well-defined outcome makes it easier for the search committee...example, "while it is possible to grow liver stem cells and liver-like mini-organs or organoids, neither...combine bioengineered scaffolds with patient stem cells to generate compatible functional livers.” Why are...Health Sciences University, did his postdoctoral fellowship with Jennifer Lippincott-Schwartz at the National... optimization of fluorescent proteins,  and live cell imaging approaches. Erik is also an avid long distance...

  13. Tips for Writing a Good Cover Letter

    Type
    Blog Post
    Published
    May 21, 2024, 1:30 p.m.
    ... grammar and spelling several times and ask someone else read your cover letter (as well as your resume...“I’d be good at that! I organized my lab data so well that I can find the results of any experiment I’...of experience in [field of research], and have excellent [list the most important skills for the position... specific examples. Instead of saying “I have excellent communication skills” try giving an example, like...

  14. Controlling for Off-target Effects with a New Genome-wide CRISPR Screen Design

    Type
    Blog Post
    Published
    Sept. 13, 2018, 1:55 p.m.
    ...off-target effects. The flaws of RNAi screens are well characterized and strategies exist to control for...delivered as a pooled library via a lentiviral vector to cell lines that either stably expressed Cas9 or that ...Hart et al, Bassik et al). The library performed well in both screens, with the growth screen having a...almost all genes involved in the production of a cell surface glycan that’s required for ricin uptake....reveals pathways underlying ricin susceptibility." Cell 152.4 (2013): 909-922. Pubmed PMID: 23394947. Pubmed...

  15. Replacing Paper: Tips for Choosing an Electronic lab Notebook

    Type
    Blog Post
    Published
    July 10, 2018, 1:08 p.m.
    ...convenient and effective, in both academic circles as well as in industry. The news is good. In their ideal...their data in one place and avoid scattering intellectual property across various software providers used...:   Does your lab need to keep paper records as well as electronic ones? It is important to know in which... it is because you need to keep paper records as well as digital, or for reporting on a project – printing... or company, site licenses might be available as well. In most cases this applies to universities. How...

  16. Plasmids 101: Gibson Assembly and Other Long-Homology Based Cloning Methods

    Type
    Blog Post
    Published
    March 1, 2016, 3:30 p.m.
    ...because of its gain in popularity. Gibson assembly is well known for allowing easy assembly of multiple linear...is immediately ready to transform into competent cells. The master mix of enzymes can be purchased from...mix for 1 hour, and then transform into competent cells. The success rate of this reaction is typically ...polymerization steps could occur. Secondly, it does not work well if the ends of the fragments have stable single ...and ordered, and are specific for each fragment as well the fragment you want next to it, so this doesn’...

  17. Scientific Reproducibility - Focusing on Solutions at the Minisymposium on Reproducibility

    Type
    Blog Post
    Published
    May 18, 2018, 7:32 p.m.
    ...directly by creating well characterised antibodies against all human extracellular proteins with community-based...are quite low (<30%, Prinz et al 2011, Begley and Ellis 2012). Recent efforts from the Reproducibility Project...Hidden “negative” data We all want to be able to tell a good story when presenting scientific findings... results. Not only is it scientifically and intellectually affirming to have a hypothesis confirmed, it...

  18. A History of Genome Engineering in Popular Culture

    Type
    Blog Post
    Published
    Feb. 25, 2020, 2:15 p.m.
    ...inspired. CAUTION: Spoilers alert! Pre-DNA Era  H.G. Wells, the father of scientific fiction, was also a trained...Asimov, Arthur C. Clarke, James Blish, and John W. Campbell. This period is termed as the golden age of SciFi... in the genome. These technologies rely on the cellular DNA repair machinery, which can lead to high frequency...1258096 . https://doi.org/10.1126/science.1258096 Gaudelli NM, Komor AC, Rees HA, Packer MS, Badran AH, Bryson... C (2000) Hugo De Vries: from the theory of intracellular pangenesis to the rediscovery of Mendel. Comptes...

  19. Lambda Red: A Homologous Recombination-based Technique for Genetic Engineering

    Type
    Blog Post
    Published
    Dec. 15, 2016, 3:57 p.m.
    ...annealing to a complementary ssDNA target in the cell.  Only Beta expression is required for recombineering...fragments, such as antibiotic resistance genes, as well as non-selectable DNA fragments, such as gene replacements...definitely the easier of the two options, but these cells are prone to mutations and will have more unintended...Central PMCID: PMC307639. 2. Daiguan Yu, Hilary M. Ellis, E-Chiang Lee, Nancy A. Jenkins, Neal G. Copeland.../Knockin Plasmids to Disrupt Genes in Mammalian Cells CRISPR Methods for Bacterial Genome Engineering ...

  20. What's New in CRISPR - Spring 2019

    Type
    Blog Post
    Published
    March 21, 2019, 12:47 p.m.
    ...temperature incompatible to genome editing in human cells. The Feng Zhang lab identified a Cas12b from Bacillus...can block Cas12a-mediated genome editing in human cells. Read the paper in Science Find the anti-CRISPR...anti-CRISPR expression plasmids CRISPR-based method for cell lineage tracing Valcyrie is a CRISPR-based marker...marker excision method for marking distinct cell clones developed by the Michalis Averof lab. It’s an alternative...
Showing: 541 - 560 of 856 results