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We narrowed to 799 results for: por

Showing: 321 - 340 of 799 results
  1. Hot Plasmids and Viral Preps - January 2021

    Type
    Blog Post
    Published
    Jan. 5, 2021, 2:15 p.m.
    ...in a protein.    Cells expressing an RFP reporter plasmid for recombinase activity were exposed ...interest. However, FACS is unable to spatially and temporally target single cells. Now, Francois St-Pierre’...way to isolate individual cells with unique spatiotemporal phenotypes from large heterogeneous cultures...introduction & resolution of two DSBs in a fluorescent reporter. Find the plasmids here Yongsub Kim’s lab engineered...
  2. Open Resources and Plasmid Tools For Studying C. elegans

    Type
    Blog Post
    Published
    July 18, 2019, 12:55 p.m.
    ...publication.   3’UTRome The 3’UTRs of mRNA are important for post-transcriptional regulation in C. elegans...combination of these methods provides unique opportunities to develop computational methods to better understand...system allows scientists to both spatially and temporally knock-down proteins of interest.  Plasmids for...intersectional strategy using a split intein for refined spatiotemporal transgene control in Caenorhabditis elegans...
  3. Overcoming the Challenges of Lentiviral Production

    Type
    Blog Post
    Published
    Aug. 7, 2018, 12:31 p.m.
    ...viral Rev gene from the packaging plasmid and incorporating it into a fourth plasmid. In addition, the promoter...third generation packaging systems, this system incorporates the Tet-off transcriptional activation system...number of harvests will all impact the titer. Importantly, optimized conditions for one transfer plasmid...producing lentivirus, it is time to titer. One important consideration when titering a lentivirus is the...
  4. Design Tips for Prime Editing

    Type
    Blog Post
    Published
    Jan. 23, 2025, 2:15 p.m.
    ...that meant rereading many of the original papers reporting various prime editing tools. These papers are ...editing application. And optimizing its sequence is important, as prime editing efficiency is affected by the...nucleotides. For longer templates, it is even more important to test different lengths and sequences, as unintended... cellular surveillance. To limit unintended incorporation of the pegRNA scaffold sequence into the genome...
  5. Hot Plasmids February 2024

    Type
    Blog Post
    Published
    Feb. 20, 2024, 2:45 p.m.
    ...difficult to genetically edit due to toxicity from electroporation or poor cellular entry with other transfection...PAGE editing with viral sgRNA targeting mCherry reporter (mChe), with co-incubation with assist peptide.... CAP-Mac also robustly expressed fluorescent reporters in green monkeys and cultured human neurons, with...slightly reduced efficacy compared to AA9-X1.1. Importantly, AAV1-X1 is not neutralized by pre-existing antibodies...
  6. Antibodies 101: Stripping and Reprobing Western Blots

    Type
    Blog Post
    Published
    Aug. 1, 2024, 1:15 p.m.
    ...directly to buffer without a re-wetting step. Important: do not dry your membrane with antibodies on it...the ones shared by Bio-Techne, ThermoFisher, or Millipore Sigma. We’ll share the basic composition of these...Alternatively, companies including ThermoFisher, Millipore Sigma, and Azure Biosystems offer pre-made stripping...your western blot several times — some researchers report up to ten cycles! Just remember that a bit of sample...
  7. Fluorescent Proteins 101: Fluorescent Protein Timers

    Type
    Blog Post
    Published
    May 4, 2017, 2:30 p.m.
    ...have been around since 2000 when Terskikh et al. reported the production of dsRed E5. This timer predictably... your experimental setup less cumbersome. 3. Portability across multiple systems Terskikh et al. showed...fluorescent timers that change color from blue to red report on cellular trafficking." Nature chemical biology...Chronology of islet differentiation revealed by temporal cell labeling." Diabetes 58.8 (2009): 1863-1868...
  8. Truncated gRNAs for Regulating Gene Expression

    Type
    Blog Post
    Published
    Jan. 10, 2017, 3:37 p.m.
    ...suffers from several limitations. Perhaps most importantly, most Cas9 orthologs are less well characterized...(1, 3). The gRNA can be further modified by incorporation of an RNA hairpin, such as the MS2 hairpin, ...with nuclease-competent Cas9. An RNA hairpin incorporated into the truncated gRNA recruits a transcriptional...target site. Benefits of truncated gRNAs Importantly, the use of truncated guides has largely been...
  9. PhD Applications After COVID

    Type
    Blog Post
    Published
    Jan. 5, 2023, 2:15 p.m.
    ...in-person portion got canceled, taking advantage of virtual meetings can still be a great opportunity to introduce...course of the last few years: recognizing the importance and benefit of remote and video communication...staff have become a new way to allow you to get important information about the campus even if you can’t...
  10. Degrading DNA with Cascade-Cas3

    Type
    Blog Post
    Published
    Feb. 11, 2025, 2:15 p.m.
    ....com.   The Cascade complex Cascade is an important component of the CRISPR Cas3 system. Most Cascade... appears to be conserved. This DNA degradation portion can be bypassed by leaving out Cas3 — a tactic ...is pulled through the Cas3. This mechanism is important for Cas3’s biggest trick. Cas3 likes to bluff,... — Cas3 will definitively degrade the targeted portion of DNA using only a single gRNA. Cas3 appears to...
  11. Viral Vectors 101: Voltage Indicators

    Type
    Blog Post
    Published
    Jan. 30, 2024, 2:15 p.m.
    ...sensors are better classified by their fluorescent reporter and the mechanism of their voltage sensing component...rhodopsin proteins were initially evolved for ion transport, not fluorescence, they are not nearly as bright...for penetrating deeper tissues depths (1 mm), important for certain applications such as in vivo tissue...rhodopsin Ace2N and mScarlet. Ace2N quenches a proportion of mScarlet's emission via FRET.  Image from ...
  12. 3D Printing Meets CRISPR Cas9

    Type
    Blog Post
    Published
    Oct. 17, 2017, 12:55 p.m.
    ...this rarely happens. But if you ever have an opportunity to read one of these structure papers – while...function in the human body?” … and probably most importantly… "how do we know this?” Over the past fifteen... nuclease active site wasn’t even close to its reported cleavage site on the complementary DNA strand....model also displays the several Lys/Ala mutations reported by Ian Slaymaker and colleagues to reduce off-...
  13. SciComm: Taking Science to Elementary and Middle Schools

    Type
    Blog Post
    Published
    July 9, 2019, 12:52 p.m.
    ...has made strong efforts in the last 5 years to incorporate more science and technology into the school curriculum...presentation to their students. I didn’t think of this opportunity as coming from networking at the time, but that...presentation because, in my opinion, it was more important to communicate the applications of CRISPR over...use it? This last section was arguably the most important, and it was a little worrisome for me since bioethics...
  14. Isolating Genomic Regions of Interest with the CRISPR System

    Type
    Blog Post
    Published
    May 30, 2019, 12:44 p.m.
    ...expression plasmid. Experimental use Since the initial report describing CRISPR-mediated purification of a specific...including Protein A, 1xFLAG, 2xAM, and HA have been reported, as well as affinity purification with an anti-Cas9...this technique, researchers have begun to make important discoveries in various biological fields. In 2015...chromatin immunoprecipitation (enChIP)." Scientific reports 3 (2013): 3171. PubMed PMID: 24201379. PubMed Central...
  15. CRISPR 101: Making a Knock-In Cell Line

    Type
    Blog Post
    Published
    Dec. 22, 2022, 2:15 p.m.
    ...typically require an exact DNA sequence to be incorporated at a precise genomic location, with little to...template for repair, which will result in the incorporation of your sequence, a.k.a a knock-in! So how does...higher HR frequencies than quiescent cells. It is important to know that HR is often mutated in cancer cells...molecule is used as the template, you need to incorporate homologous sequence to the right and/or left ...
  16. An Introduction to Adenovirus

    Type
    Blog Post
    Published
    Oct. 3, 2023, 1:15 p.m.
    ...toward saying that host-virus interactions are an important part of the biology of adenoviruses (AdVs). AdVs... Although host-virus interactions likely seem important for any virus, we will see that they are very ...(Tollefson et al., 1996) – cell death has been reported when using vectors, and immune inactivation of...Adenovirus Tales: From the Cell Surface to the Nuclear Pore Complex. PLoS Pathog, 11(6), e1004821. https://doi.org...
  17. Tips for arabidopsis transformation

    Type
    Blog Post
    Published
    Oct. 25, 2018, 1:23 p.m.
    ...analyzable transgenic in hand, so it is pretty important to get this process right the first time. I’ll...to transform when they begin flowering. It is important to perform the transformation not long after flowering...cut off the primary inflorescence (the flowering portion of the plant) near the base of the stem. This buys...s time to select for transformants. It’s very important to select multiple T1 plants from each transformation...
  18. Better Dyeing Through Chemistry & Small Molecule Fluorophores

    Type
    Blog Post
    Published
    Sept. 8, 2016, 2:30 p.m.
    ...transcyclooctene–tetrazine)– Nonnatural amino acids can be incorporated into a protein structure and then used with ...attach a fluorophore at the specific site of incorporation (Figure 1d). Fluorogen activating proteins...excess dye from the sample, which is particularly important for samples where washing is difficult, such as...inside living cells. We recently discovered that incorporation of four-membered azetidine rings could substantially...
  19. Interview: Hodaka Fujii on enChIP, New CRISPR Tools, and More

    Type
    Blog Post
    Published
    Dec. 2, 2014, 7:23 p.m.
    ..., as well as the Editorial Board of Scientific Reports and PLoS One. Additionally, you are involved in...is being involved in the scientific community important to you and your lab? Fujii: I first knew Addgene...interact with each other, which is critically important, especially for young researchers. It was a really...technologies. Majority of them are eager to just "import" technologies developed abroad, so it's a little...
  20. Deciphering the Mysteries of Behavior with Viral Vectors

    Type
    Blog Post
    Published
    Sept. 17, 2019, 12:40 p.m.
    ...Geneva (Creed et al., 2015) used optogenetics to support these existing findings by controlling neurons ... animal is placed into a tank of water with no support from below or anything to hold on to. In general..., 2007). A study from 2014 (Bahi et al., 2014) supports the relevance of BDNF in depression. The researchers...of viral vectors to fine tune gene expression temporally and spatially, exciting times are ahead for behavioral...
Showing: 321 - 340 of 799 results