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Showing: 201 - 220 of 872 results
  1. Visualizing Genomic Loci with CRISPR-Sirius

    Type
    Blog Post
    Published
    April 18, 2019, 12:53 p.m.
    ...cells (Ma et al., 2015). To do so, they fused a different fluorescent protein to various Cas9 variants, ...either MS2, PP7, or boxB) that each recruits a different fluorescent protein (BFP, GFP, or RFP) to the ...green, or red readout. But gRNAs containing two different hairpins will produce a secondary color (yellow...allows for simultaneous visualization of sites on different chromosomes in living cells, the system requires...they chose an intergenic DNA region (IDR3) as a reference loci to measure inter-locus distances of loci ...sgRNA-Sirius-4X(MS2-PP7) showing recruitment of different fluorescent proteins. (B) CRISPR-Sirius used to...try. Find CRISPR-Sirius plasmids at Addgene. References Ma, Hanhui et al. “Multicolor CRISPR labeling...
  2. Addgene Depositors Get More Citations

    Type
    Blog Post
    Published
    April 20, 2017, 2:30 p.m.
    ...deposits to Addgene.  Insomuch as different journals represent different research areas, target readers,... would also be great to know how much of that difference in citations is caused by depositing a plasmid...wouldn’t be causal. To control for a number of differences between those papers with Addgene plasmids and..., so our estimates now take into account any differences in citations due to the journal of publication... the same journals! So, is all this remaining difference caused by depositing the related plasmids with...it is still suggestive that we see such a big difference in citations after controlling for journal. And...Visit his website at: http://www.neil-t.com/ .   References 1. Cong, Le, et al. "Multiplex genome engineering...
  3. R Bodies: Membrane-Rupturing Microscopic Tools

    Type
    Blog Post
    Published
    April 14, 2016, 2:30 p.m.
    ...not exert force by dynamically rearranging into different polymerization states. Instead, R body monomers...natural context or 3) be modified to extend at different pH levels. Domesticating R bodies We have shown...that R bodies can function to break membranes of different organisms. And finally, we demonstrated that R... can be “tuned” with mutagenesis to extend at different pH values. Using a screen that takes advantage... fact that coiled R-bodies sediment in certain buffers but remain soluble when extended (see figure to...region produced R-bodies that could extend at different pHs. Plasmids expressing expressing both wild ...School. Follow her on Twitter @jessicapolka.     References 1. Kanabrocki, J. A., Quackenbush, R. L. & Pond...
  4. Components of CRISPR/Cas9

    Type
    Blog Post
    Published
    Feb. 2, 2016, 5 p.m.
    ... labs (see table below) have different PAM requirements. These different PAM requirements allow researchers...discovered all the time and the requirements of these different systems can vary (for example, xCas9 is a variant...neither bind nor cut the target. Cas9 homologs from different organisms or Cas9 mutants developed in a variety...researchers to target many different genomic loci. 3. Cleave target DNA resulting in a double-strand break...). Throughout the remainder of this post, we’ll refer to this RNA complex as the “gRNA”. The gRNA contains...choice (see tips for designing a gRNA) and their prefered Cas9 variant (with the appropriate PAM sequence...sequence.   Now that you know about all the different components of CRISPR, It's time to start thinking...
  5. Advice for Scientists Starting a Lab

    Type
    Blog Post
    Published
    April 29, 2014, 1:43 p.m.
    ...student or postdoc may require different types of mentoring. Some mentees prefer lots of advice and micromanagement...an effective working environment? Tom Ellis was offered a start-up package that included a postdoc and ...recommends to seek out advice from seasoned PIs who can offer support. “It is an exciting and emotional time!”...recruit people who are nice as top criterion and offers advice to not rush when growing your lab. “One ... lab and safety manager, researcher, adviser, conference planner, and writer.” Connie Cepko was surprised...impossible' or useless.” How do you run your lab differently or the same as your adviser did? Tom Ellis merges...online. The Howard Hughes Medical Institute (HHMI) offers some great resources for Early Career Scientists...
  6. 22 Hot Plasmid Technologies from 2014

    Type
    Blog Post
    Published
    Jan. 6, 2015, 5:21 p.m.
    ...libraries expand upon the CRISPR family of plasmids by offering a lentivirus-based mechanism for sgRNA delivery... microscope 8 hours post-pulse. The distinct differences of red-to-green fluorescence in individual mitochondria...mitochondria in the networks marks the differences in import of newly synthesized (green) protein and ...five previously unknown channelrhodopsins from different species: Stigleoclonium helveticum (ShChR), Chlamydomonas...advances, the Ting lab’s first-generation APEX suffered from inconsistent activity levels and limited ...cassette. They validated this methodology in two differents types of bacteria (E. coli and Salmonella typhimurium...naturally-occurring variations in the TALE repeat, referred to as “non-RVD variations”, were found to improve...
  7. Viral Vectors 101: AAV Serotypes and Tissue Tropism

    Type
    Blog Post
    Published
    Jan. 16, 2025, 2:15 p.m.
    ...2025 and updated in August 2025. References and resources References Ahuja, V., Jeyabalan, S., & Tzanakakis...multiple coreceptors, which may be present on different cell types and sometimes vary between target species... numbers separated by a slash: the first number refers to the genome, and the second number to the capsid...AAV8, AAV9 In addition to tissue tropism, different AAV serotypes can be useful to evade neutralizing... (AAV) Serotype 9 Provides Global Cardiac Gene Transfer Superior to AAV1, AAV6, AAV7, and AAV8 in the ...Hemagglutination and Efficient Transduction but Differ in Sialic Acid Linkage Specificity. Journal of ... Dudman, J. T., Hantman, A. W., Looger, L. L., Schaffer, D. V., & Karpova, A. Y. (2016). A Designer AAV...
  8. Plasmids 101: Dimers and Multimers

    Type
    Blog Post
    Published
    Jan. 10, 2023, 2:15 p.m.
    ...plasmid sample has a mix of different-sized multimers, you may even see different sets of bands on the gel... us at [email protected]. Resources and References References Bacolla A, Wang G, Jain A, Chuzhanova NA...Nanopore sequencing or PacBio SMRT sequencing, (which differ from the short-read Illumina MiSeq sequencing that...multimerization has been observed to occur in many different kinds of plasmids, taking many forms (Levene, ...from BiteSize Bio for more information on how different plasmid conformations run on an agarose gel). ...low metabolic burden seed-stock and fed-batch fermentation processes. Biotechnol Bioeng. 2009 Aug 15;103...Plasmids 101 eBookAddgene's Molecular Biology ReferenceAddgene's Restriction Digest Protocol More resources...
  9. A Control for All Seasons

    Type
    Blog Post
    Published
    Oct. 5, 2023, 1:15 p.m.
    ...check out these blog posts.       References and resources References Burry, Richard W. “Controls for ..., you will have a better understanding of the different types of controls and be able to use this information...treatment versus and which are likely just natural differences between individuals. Recommended controls for...extremely bright, then the noise likely will not interfere. If the fluorescence from your target is weak,...fluorescence of the sample, or autofluorescence, also interferes with data interpretation and can be addressed....  Western blotting When comparing samples in different lanes in a western blot, you’ll need to include...equal loading across samples and equal protein transfer from the gel to the membrane. A loading control...
  10. AAV Q&A with Tim Miles

    Type
    Blog Post
    Published
    May 17, 2022, 1:15 p.m.
    ... the best method for titering AAV? Is there a difference in titer results between ddPCR and qPCR? A: At...curve and somewhat more reliable quantitation. Differences in sample treatment across methods can affect...  Q5: What is the efficiency of co-delivering different AAV particles into the same cells in vivo? A: ...give some insight into codelivery efficiency at different doses. See Figure 4 in this paper for more information...very high dosage. Any thoughts?  A: The question refers to this paper. AAV can be safely concentrated to...IACUC committee.  Animal Models Q9: Is there any difference in transfection efficiency when delivering these...injections? A: We have not observed any appreciable difference in AAV transduction efficiency between retro-...
  11. Choosing a CRISPR Nuclease: Site Accessibility, Specificity, and Sensitivity

    Type
    Blog Post
    Published
    Nov. 5, 2019, 1:28 p.m.
    ...isolated from different organisms and engineered Cas9 variants that were able to target different PAM sites...activity of Sniper-Cas9 to different Cas9 variants using gRNAs of different lengths (Lee et al., 2018)...found in other species can recognize a variety of different PAM sequences. Researchers have also used experimental...activity relative to Cas9. Since then, the Joung and Jennifer Doudna Labs used targeted mutagenesis to generate...relative to eSpCas9 and SpCas9-HF across a number of different sites. Using a combined random and directed mutagenesis...choosing a Cas protein for your experiment. References Hu, Johnny H., et al. "Evolved Cas9 variants ...
  12. Viral Vectors 101: An Introduction to AAV

    Type
    Blog Post
    Published
    May 30, 2023, 1:15 p.m.
    ... the retina using viral tools.   References and Resources References Pillay, S., et al. "An essential ...and can be specifically targeted to a number of different cell types even when systemically administered...researchers speak about AAVs, they are actually referring to rAAVs. ITRs and serotypes The names of AAVs...ITRs, which comes from the AAV2 serotype. Serotype refers to the type of capsid (outer protein shell) used...capsids allow the virus to infect (or not infect) different cell types with some specificity. There are a ...will lose expression quicker than terminally differentiated, non-dividing cells.   Figure 2: ...subretinal injections of neonatal mouse eyes at two different ages." Retinal Degenerative Diseases. Springer...
  13. 5 Steps to Writing Grant Proposals for Grad Students

    Type
    Blog Post
    Published
    Feb. 11, 2014, 12:13 p.m.
    ... and offer educated insight into what the desirable features of future technologies should offer in order... funding towards my PhD project. Grant writing offers various skill building opportunities that graduate... to a given problem, Empowers the writer, And offers insight into a career as a Professor. Step ...Challenge motivates the request for application and offers an opportunity to summarize the limitations of ...potential pitfalls of the intended protocol, and offers alternative solutions to demonstrate thorough thought...standard “Our work will empower the world” tone that differs from publication writing and may be challenging...and exposure, here are some additional links that offer more advice on writing a grant proposal. "Murder...
  14. Popular Retroviral Vectors and Their Uses in Scientific Research

    Type
    Blog Post
    Published
    July 24, 2018, 12:51 p.m.
    ...tools for genetic engineering. There are several different viral vectors to choose from, for example gamma-retrovirus...separate envelope and packaging plasmids as well as transfer plasmids. This concept ensures safe handling and...isoforms of these proteins, as well as the viral LTRs differ. Therefore, lentiviral and gamma-retroviral packaging...Gamma-retroviral vectors Popular gamma-Retroviral Transfer Vectors   Expressing your gene of interest pMKO...expressing RNAi pMKO.1 puro (Plasmid #8452) RNA interference (RNAi) is an RNA-mediated gene silencing mechanism...interest. Lentiviral vectors Popular Lentiviral Transfer Vectors    Expressing your gene of interest pLenti-puro... in phenotype. The plasmid comes with a 1.9 kb stuffer sequence that can be removed and replaced with ...
  15. BeHeard Award 2018: Diseases of Glycosylation, Arginine Mutagenesis, & Neural Development

    Type
    Blog Post
    Published
    June 26, 2018, 1:33 p.m.
    ...pathological conditions. References: Monticelli, M., Ferro, T., Jaeken, J., dos Reis Ferreira, V. & Videira, P...GdnHCl to patients suffering from rare diseases arising from Arg mutations. References: Baldwin, Enoch,...life-threatening consequences. They result from defects in different steps in the glycan biosynthetic pathway and have...recently reviewed and reported that PMM2-CDG patients suffer from immunological defects. These are characterized... discovered that the in vitro functions of 11 different Arg-mutant enzymes could be rescued by supplementation...
  16. Design Tips for Prime Editing

    Type
    Blog Post
    Published
    Jan. 23, 2025, 2:15 p.m.
    ...Figure 9: epegRNAs (left) and PE7 (right) offer two different strategies for protecting pegRNAs from degradation...software in the comments below! References and Resources References Anzalone, A. V., Randolph, P. B.,... These tips are a great starting point: Test different lengths of the primer binding site, starting with...still be optimized (Anzalone et al., 2019). Test different lengths of reverse transcriptase template, starting... templates, it is even more important to test different lengths and sequences, as unintended secondary...specific prime editing tools There are a lot of different prime editing tools to choose from, and many have...
  17. Getting Started with Behavioral Neuroscience: Standardizing Experiments and Using Viral Vectors

    Type
    Blog Post
    Published
    Aug. 20, 2019, 12:31 p.m.
    ... can only carry around 4.5 kb. Different AAV serotypes have preferences for specific cell types. For example...behavioral studies. References Stoica L, Ahmed SS, Gao G, Sena-Esteves M. Gene transfer to the CNS using ...the experiment. Handling can however negatively interfere with certain tests and is not always recommended...end of an experiment series would behave very differently (distracted) from the ones tested at the beginning...Experiment Behavioral scoring systems (experimenter differences in behavior observation); experimenter blinding...injections. AAV particles are smaller and may be preferable if wider diffusion is required. On the other ...
  18. Plasmids 101: Modular Cloning

    Type
    Blog Post
    Published
    March 12, 2024, 1:15 p.m.
    ...you can quickly test that part in different contexts and plug different characterized parts into a TU! What...works on genetic code expansion.  References and Resources References Weber E, Engler C, Gruetzner R, Werner...cloning is MoClo. That’s because MoClo specifically refers to the organizational schema described above, and...nature of MoClo allows users to test a variety of different orientations and components within the system ... a great tool to deliver many combinations of different candidate genes at once to identify the best cocktail...and gene editing that use MoClo, for which many different MoClo kits have already been established. Keep...
  19. A Look Back at One Year of Plasmid Sharing for COVID-19 Research

    Type
    Blog Post
    Published
    July 12, 2021, 1:15 p.m.
    ...never been so important.     References and resources References Biering SB,  Sarnik SA, Wang E, Zengel... study the mechanisms of infection and how to interfere with it in a BSL-2 lab instead of a BSL-3 lab....for generating pseudovirus and those encoding different spike protein variants that can be incorporated...vaccinated individuals are at neutralizing the different SARS-CoV-2 spike variants. You can learn more ...widely used qPCR and antigen tests.  Labs including Jennifer Doudna’s lab and Feng Zheng’s lab had already ...head over to this blog post that highlights five different methods to use CRISPR to detect SARS-CoV-2. Other... Jennifer Tsang...
  20. Science communication: 9 Strategies to Get Your Foot in the Door

    Type
    Blog Post
    Published
    Sept. 27, 2017, 1:23 p.m.
    ...publish a new blog post, and live tweet from conferences. Find more tips on using social media as a scientist...for example at an anniversary or for Girls’ Day, offer guided tours to show people where the lab magic ...communicate science. You can draw live sketches at conferences like @ATJCagan or create agar art with colorful...Association, EUSJA or ABSW for helpful tips and job offers. If you live in the UK, join the SciComm mailing...10 days. And the Society of Experimental Biology offers press internships to cover their annual meetings...College London. Similarly, the University of Cardiff offers an MSc in Science, Media and Communication. In ...Santa Cruz, MIT, UW Madison, and Boston University offer science communication programs. Others, like Northwestern...
Showing: 201 - 220 of 872 results