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We narrowed to 68 results for: des.1

Showing: 1 - 20 of 68 results
  1. How to Design Your gRNA for CRISPR Genome Editing

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    Blog Post
    ...non-functional allele. For a 1 kilobase gene, since potential target sites occur ~1 in every 8 nucleotides,...Vazquez F, Parts L, Boehm JS, Golub TR, Hahn WC, Root DE, Garnett MJ, Tsherniak A, Iorio F (2019) Agreement...770–788 . https://doi.org/10.1038/s41576-018-0059-1 Veres A, Gosis BS, Ding Q, Collins R, Ragavendran ...j.stem.2014.04.020 Yang L, Guell M, Byrne S, Yang JL, De Los Angeles A, Mali P, Aach J, Kim-Kiselak C, Briggs...
  2. QC Sequencing Technologies at Addgene

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    Blog Post
    ...and recorded as a DNA strand is synthesized (Figure 1). Sanger sequencing requires four separate reactions...trace” file to determine the sequence. Figure 1: Overview of Sanger sequencing technology. Created... apc, and NOVOPlasty. The pipeline contains three de novo assemblers, so if one fails to generate a circular...manually checks and assembles the raw data.  Addgene’s de novo assembly pipeline usually generates the correct...raw FASTQ reads are used for either guided or new de novo assembly. If a full anticipated sequence is ... doesn’t resolve the assembly issues, we can redo de novo assembly within Geneious.  While this may seem...steps that gave us our initial incorrect assembly, de novo assembly within Geneious gives us additional...
  3. Bricking Science: Portraying Scientific Reality Through LEGO

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    Blog Post
    ...thanks to our guest blogger, Dalila Cunha de Oliveira. Dalila Cunha de Oliveira is a PhD student in Hematology...post was contributed by guest blogger, Dalila Cunha de Oliveira. Bricking Science is an idea built, literally...motion videos is very time consuming; producing a 1-2 minute video takes an entire week of our free time...
  4. CRISPR Protocol for Genomic Deletions in Mammalian Cell Lines [Video]

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    Blog Post
    ... nucleases. Genome Res. 2010 Jan;20(1):81-89. PubMed. 2. Bauer DE, Kamran SC, Lessard S, Xu J, Fujiwara...harness non-homologous end joining repair (NHEJ) [1]. Our initial work was with TALENs, in collaboration...from the University of Pennsylvania.     References: 1. Lee HJ, Kim E, Kim JS. Targeted chromosomal deletions...11;342(6155):253-257. PubMed. 3. Canver MC, Bauer DE, Dass A, Yien YY, Chung J, Masuda T, et al. Characterization...
  5. Adeno Associated Virus (AAV) for Cell and Gene Therapy

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    Blog Post
    ...thanks to our guest blogger, Harshana S De Silva Feelixge. Harshana S De Silva Feelixge is a researcher whose... post was contributed by guest blogger Harshana S De Silva Feelixge. Gene therapy technologies hold ...reconstitution with a normally functioning T cell repertoire (1). In this example, it was possible to transduce the...significant improvements in vision over a period of 1 to 3 years – the treatment appears to be both safe...research and science communication.     References 1. Gaspar, H. B., S. Howe, and A. J. Thrasher. "Gene...
  6. A Look at Addgene's QC Process

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    Blog Post
    ...experiments, to false conclusions.  Figure 1. Potential points where DNA errors can be introduced...are assembled into a circularized plasmid sequence de novo (without the use of a reference sequence) using...for the plasmid, QC scientists will first align the de novo NGS result to the deposited sequence(s) in SnapGene...sequence, if one was provided. If no reference is given, de novo assembly in Geneious can still provide additional...
  7. CRISPR Plasmids - Cascade-Cas3

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    Collection
    ...Cascade ( C RISPR- as sociated c omplex for a ntiviral de fense) complex, comprised of a combination of Cas5...endogenous repair mechanisms. Cas3 belongs to the Class 1 family of CRISPR systems, the most abundant type found...bacteria and archaea. While abundant in nature, Class 1 systems are largely underutilized compared to their...still separate the individual Cas components. Figure 1: Overview of Cascade-Cas3 mechanism. Created with ...
  8. Crowdfight, a Platform to Boost Scientific Collaboration During COVID-19 and Beyond

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    Blog Post
    ...postdoctoral researcher at Instituto de Investigación Biomédica de Salamanca (IBSAL) and one of the coordinators...prompted us to create Crowdfight.   Figure 1: Crowdfight is a community of scientists who believe...collaborator for a specific task, Crowdfight will: 1) Find an expert with the right skill; 2) Confirm the...
  9. FLEx Technology and Optogenetics: Flipping the switch on gene expression with high spatial and temporal resolution

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    Blog Post
    ...halorhodopsins to silence neuronal activity (Figure 1) (Wiegert et al., 2017). How to get an opsin into... rather than aversive stimuli. In a recent paper, de Jong and colleagues used rAAV FLEx-based vectors ...brain regions associated with motivated behaviors (de Jong et al., 2019). FLEx vectors represent, therefore...developmental biology at the École Normale Supérieure de Lyon. Her love for educational science led her to...neuroscience 8.9 (2005): 1263. PubMed PMID: 16116447. de Jong, Johannes W., et al. "A neural circuit mechanism...
  10. Visualizing Protein Turnover In Situ

    Type
    Blog Post
    ...unpublished, that allowed the visualization of specific de novo synthesized proteins in situ. I was privileged..., 2010) and allows the visualization of the whole de novo synthesized proteome. However, FUNCAT alone ... contact her at [email protected]. References 1. Dieterich, Daniela C., et al. "In situ visualization...
  11. CRISPR Plasmids - CRISPR Transposases (CAST)

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    ...Cascade ( C RISPR- as sociated c omplex for a ntiviral de fense), and four transposase components (TnsA, TnsB...some mammalian systems have been developed. Figure 1: Overview of the basic CAST mechanism. Created with...
  12. PRIDICT: Predicting Efficiencies of Prime Editing Guide RNAs

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    Blog Post
    ...References Akhtar, W., De Jong, J., Pindyurin, A. V., Pagie, L., Meuleman, W., De Ridder, J., Berns, A....evaluate all of the pegRNAs in one experiment (Figure 1). Through this method, we found key determinants of...diverse edit types and cell contexts. Figure 1: High-throughput screening of pegRNA efficiency and...Article 3. https://doi.org/10.1038/s41576-022-00541-1 Kim, H. K., Kim, Y., Lee, S., Min, S., Bae, J. Y.,...DNA repair determinants. Nature Biotechnology 2023, 1–11. https://doi.org/10.1038/s41587-023-01678-y Li,...editing systems in multiple cell types. Cell, 186, 1–17. https://doi.org/10.1016/j.cell.2023.03.034 Resources...
  13. Celebrating the 2024 Nobel Prize in Chemistry

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    Blog Post
    ...existing protein structures, called Top7. Figure 1: Top7 receives the Nobel Prize. Created with BioRender.com...from the Baker Lab and others. The introduction of de novo designed proteins also helped researchers understand...
  14. Save Time with Transient Plant Leaf Transformations

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    Blog Post
    ... medicinal plant, the Catharanthus roseus (Figure 1). The ultimate goal of our research is to enhance ... which one you choose depends on your experiment (de Ruijter et al., 2003). For example, we did our initial...Humana Press, 2009. 41-50. PubMed PMID: 19183892. De Ruijter, N. C. A., et al. "Evaluation and comparison...activities in seed plants." Plant cell reports 9.1 (1990): 1-5. PubMed PMID: 24226366. Lee, Min Woo, and Yinong...
  15. Plasmids 101: Expressing shRNAs for RNA Interference

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    Blog Post
    ...vectors. BMC Biotechnology, 6(1). https://doi.org/10.1186/1472-6750-6-1 O’Brien, J., Hayder, H., Zayed...endogenous miRNA machinery for processing (Figure 1). The shRNA sequence is transcribed to produce hairpin... recruits other silencing proteins. Figure 1: Comparison of shRNA and miRNA processing and silencing...,"rgb":"rgb(0, 82, 169)","rgba":"rgba(0, 82, 169, 1)"},"button_font":{"size":19,"styles":{"font-family...:"rgb(255, 255, 255)","rgba":"rgba(255, 255, 255, 1)"}}{% endraw %}{% end_module_attribute %}{% module_attribute...297. https://doi.org/10.1016/s0092-8674(04)00045-5 De Rie, D., Abugessaisa, I., Alam, T., Arner, E., Arner...Gebhard, C., Goldowitz, D., Guhl, S., Ha, T. J., . . . De Hoon, M. J. L. (2017). An integrated expression atlas...
  16. Botman-Teusink Yeast FP Collection

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    ...Link opens in a new window) References Botman, D., de Groot, D. H., Schmidt, P., Goedhart, J., Teusink,...fluorescent proteins in budding yeast. Sci Rep, 9 (1):2234. https://doi.org/10.1038/s41598-019-38913-z ...
  17. Viral Vectors 101: AAV Variables That Matter

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    Blog Post
    ...vectors. PLoS ONE, 9(8), 1–10. https://doi.org/10.1371/journal.pone.0106472 de Backer, M. W. A., Brans,...promoters. Gene Therapy, 28(1–2), 56–74. https://doi.org/10.1038/s41434-020-0169-1 Oh, J. Y., Han, J. H., ...the selection process. Factors to think about are (1) your experimental conditions and measurements; (2...serotype, promoter, and titer.     Figure 1: Flow chart depicting a) steps for selecting serotypes...injection site than AAV2 (Watakabe et al., 2015) (Figure 1). You’ll need to consider if such properties could...AAV.  Figure 2: Comparison of AAV serotypes 1, 2, 5, 6, 8, and 9 in two different brain regions....Image modified from Aschauer et al., 2013, Figure 1. Used under Creative Commons Attribution License. ...
  18. Neurodegeneration Plasmid Collection

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    Collection
    ...Fawzi 127194 RP1B FUS 1-163 QQ4xSS #1 FUS His T7 ALS Nicolas Fawzi 127195 RP1B FUS 1-163 QQ4xSS #2 FUS His... type 1 scFv [L24/1] ITPR1 CMV Spinocerebellar ataxia James Trimmer 206790 IP3 receptor, type 1 scFv [... pLKO.1 mTagBFP2 rat DJ-1 shRNA PARK7 U6 Parkinson's Timothy Ryan 223563 pmCherry-Synaptojanin-1-145 SYNJ1...Cookson 13321 pGEX5X.1 PINK1 WT PINK1 GST tac Parkinson's Mark Cookson 13322 pGEX5X.1 PINK1 KD PINK1 GST...pcDNA3-FLAG-Synaptojanin 1-145 SYNJ1 Flag CMV Parkinsonism, early-onset Pietro De Camilli 22292 pcDNA3-FLAG-Synaptojanin...-FLAG-Synaptojanin 1-170 SYNJ1 Flag CMV Parkinsonism, early-onset Pietro De Camilli 22293 pEGFPC1-FLAG-Synaptojanin...FLAG-Synaptojanin 1-145 SYNJ1 GFP, Flag CMV Parkinsonism, early-onset Pietro De Camilli 22294 pEGFPC1-FLAG-Synaptojanin...
  19. Rett Syndrome: A History of Research and Therapeutic Outlooks

    Type
    Blog Post
    ...in transcriptional repression, activity dependent de-repression, chromatin remodeling, gene activation...therapies to clinical trials in 2021. Figure 1: Schematic of the human MECP2 locus and MECP2 protein...Tillotson R, Selfridge J, Koerner MV, Gadalla KKE, Guy J, De Sousa D, Hector RD, Cobb SR, Bird A (2017) Radically...
  20. Tips for a 1st Time CRISPR User (by a 1st Time CRISPR User)

    Type
    Blog Post
    ...The entire process from start to finish took about 1 month. Cas9 delivery Overview: Before any editing...lentiviral vector, lentiCas9-Blast (52962-LV, Figure 1). Some cells were then used to generate a monoclonal...rest of the cells were frozen down. Figure 1: Cas9 expression in A549 cells transduced with lentiCas9...Western blotting is a loading control. Pro-Tips: 1. It’s all about the MOI. In retrospect I used a way... pool had wildly variable Cas9 expression (Figure 1). This is a problem because Cas9 expression affects...cells were transduced (MOI = 37) and selected with 1 µg/mL blasticidin for 9 days. Single cells from a ... and from individual cell expansions (monoclonal, 1-5). Wild-type A549 cells do not express CAs9 and are...
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