... and education 3:49-4:35 : Kingdom on opportunities to go into science in Malawi 4:36-5:46 : How Kingdom...more about Dr. Kwapata’s current work and thoughts on the need for more scientific collaboration across... connecting us with him. Learn more about our recent collaboration with Seeding Labs here. Special thanks...received an award from nonprofit and Addgene collaborator, Seeding Labs, for lab equipment that will help...Addgene Podcast.     Additional Resources Learn about Seeding Lab's Work with Lilongwe Univeristy of Agriculture...How You Can Contribute to the Addgene Blog Learn about Addgene Plasmid Donations for Rare Disease Research...

... min) Hot Plasmids podcast that introduces you to 4 new plasmid technologies from one of our recent newsletters...The Human Kinase Domain Constructs Kit Hot Plasmid 4: The Yeast Prototrophy Kit from Markus Ralser's Lab...this new way to learn about plasmid technologies. If you have any comments about the Hot Plasmids Podcast...with these efforts, you’ll find new ways to learn about science careers, lab protocols, and, of course, ...each one, plug in some headphones to learn more about the latest and greatest plasmids through this podcast...

...competent for excision, but defective for integration [4], the donor cassette is removed leaving behind no ... with the wild-type piggyBac™ transposase enzyme [4]. By using a donor plasmid containing a negative selection...Methods Mol. Biol., Apr. 2016. PubMed PMID: 27075976. 4.  X. Li, E. R. Burnight, A. L. Cooney, N. Malani, ...target locus modifications.,” Mol. Ther., vol. 18, no. 4, pp. 743–53, Apr. 2010. PubMed PMID: 20068556. PubMed... retroviral insertion sites.,” Nat. Protoc., vol. 4, no. 5, pp. 789–98, Jan. 2009. PubMed PMID: 19528954...transposase itself, please read on to learn more about this exciting technology from the folks at Transposagen...Additional Resources on the Addgene Blog Learn about the Sleeping Beauty Transposase Read up on Genome...

...Depending on the protocol, samples can be expanded to ~4-10x linearly! This expansion physically separates ... cases, labels are added after this stage.  Step 4: Expand! (Typically by just adding water.)  Step 5...quite so much. Created with biorender.com Steps 2-4 are not found in standard IHC/ICC protocols and you...16:33–41. https://doi.org/10.1038/s41592-018-0219-4  Yu C-C (Jay), Barry NC, Wassie AT, et al (2020) Expansion...applications and there’s just not much to be done about that.  Second, expansion introduces distortions....

...under the highest biosafety level conditions, BSL-4. In this regard, minigenome systems, such as the one...the EBOV replication cycle under BSL-2 conditions (4). What is a minigenome?   The EBOV genome is a nonsegmented... ratios result in significantly reduced activity (4). To ensure that you end up with appropriate plasmid...PMID: 21699921. PubMed Central PMCID: PMC3586226. 4. Mühlberger, Elke, et al. "Comparison of the transcription...Microbiology, National Emerging Infectious Disease Laboratories.  She is interested in genome replication and...

... single gene KO and pooled screening approaches; (4) encodes a Thy1.1 selectable marker to facilitate ...Framework for in vivo T cell screens. J Exp Med, 221(4), e20230699. https://doi.org/10.1084/jem.20230699....faithfully model the complex spatial, cellular, metabolic, and cytokine microenvironment that exists in ...LaFleur is a Postdoctoral Fellow in Arlene Sharpe’s laboratory at Harvard Medical School and is interested in...

...codelivery efficiency at different doses. See Figure 4 in this paper for more information.   Q6 & Q7: We ...organism such as mice, e.g. first at 2 months, then at 4 months? Or do antibodies become a prohibitive problem...: CAP-B10 and CAP-B22 were tested individually in 4 marmosets each. Brain potency for each individual ...labeling in the brain with PHP.eB: A recent CLOVER collaboration injected 1e+13 vg per mouse to get phenotypes...coverage. Please see this paper for more information about PHP.eB brain labeling at various doses.  As always...

...small molecule derivative of GFP called 3,5-difluoro-4-hydroxybenzylidene imidazolinone (DFHBI). DFHBI is... ligands by exponential enrichment (SELEX). After 4-6 rounds of SELEX, the RNA pool was cloned into bacterial... dimer activates the fluorescence of 3,5-difluoro-4-hydroxybenzylidene-imidazolinone-2-oxime (DFHO), a...fluorescence with light-up aptamers." Frontiers in chemistry 4 (2016): 29. PubMed PMID: 27446908. PubMed Central ...occurring regions in the 5’ UTR of mRNA that bind metabolites, leading to conformational changes that control...Spinach-based riboswitches can be used as stable metabolite sensors for imaging in live cells. By placing...placing Spinach downstream of a riboswitch, metabolite binding to the riboswitch instead activates Spinach ...

... formation of plasmid multimers. Cell. 1976 Dec;9(4 PT 2):707-16. doi: 10.1016/0092-8674(76)90134-3. PMID...in bacteria. Proc Natl Acad Sci U S A. 1968 Dec;61(4):1406-13. doi: 10.1073/pnas.61.4.1406. PMID: 4884687...capillary gel electrophoresis. BioProcess Int. 2006 Sep;4(8):38-40. Summers D. Timing, self-control and a sense...plasmid monomerization and stability. Cell. 1984 Apr;36(4):1097-103. doi: 10.1016/0092-8674(84)90060-6. PMID...upstream production. Biotechnol Adv. 2009 Jul-Aug;27(4):353-70. doi: 10.1016/j.biotechadv.2009.02.003. Epub...colonies using one of the detection methods described above can help ensure the isolation of the monomeric version...plasmid DNA production platform incorporating low metabolic burden seed-stock and fed-batch fermentation processes...

...expression. When using these constructs to tag the GOLG-4 protein, mScarlet3 was 50% brighter than the original... Figure 3: Expression of fluorophore-tagged GOLG-4 through eight stages of embryonic development. Representative...knockouts limits further cutting and errors (Figure 4). This work marks the first use of CRISPR/Cas9 in ...pathogenicity, and resistance mechanisms.     Figure 4. Overview of the dual-plasmid CRISPR/Cas9 workflow...more feasible, the lab of Alessio Ciulli and collaborators developed CRBNmidi (Kroupova et al., 2024). ...stepwise, achieving mutation efficiencies of up to about 9% of colonies (102 to 104 times higher than traditional...Bentley Time to get flexible! Alex Dunn’s lab and collaborators have deposited a new mechanosensor called PILATeS...

...2in1 Plasmid Toolkit consists of 4 plasmid toolkits generated with 4 different, binary 2in1 vectors with...combinations of tag orientation (NN, NC, CN, and CC). The 4 kits also contain the two entry vectors pUC-L1L4 and... plasmids you need. If you'd ever like to write about a recent plasmid deposit please sign up here.    ...were observed binding telomeric DNA. What’s great about CASANOVA? It works with the S. pyogenes Cas9, dCas9...

...Neuroscience. 2022. https://doi.org/10.1038/s41593-021-00969-4.   Increased neuronal expression using Cre-mediated...Neuroscience. 2022. https://doi.org/10.1038/s41593-021-00969-4.   dCas9-SSAP with reduced error rate for long sequence...to be deconvoluted using freeware.    Fig. 4: Images of 12 visual barcodes in use in A375 cell ... plasmids you need. If you'd ever like to write about a recent plasmid deposit please sign up here. ...

...board, Jacqueline N Crawley  [et al]. 2013;4(435):4 35 1-4  20. doi: 10.1002/0471142301.ns0435s65. PMID: ... different variants [1, 2, 4]. And, of course, the best way to learn about the many different opsin variants...animal models. Nature reviews Neuroscience. 2012;13(4):251-66. doi: 10.1038/nrn3171. PMID: 22430017. 2. ....2011.10.003.  PMID: 22055387;  PMCID: PMC3225502. 4. Mattis J, et al. Principles for applying optogenetic... electrophysiology data too but that’s what collaborators are for). For example, if you study neural pathways...intended brain region and/or cell type (I’ll talk more about this in detail later). But what are the different...of your opsin, but keep you eyes open for posts about the next steps in the coming weeks. Expression of...

...vacuole and emit red and green fluorescence. After 4 hours of nitrogen starvation to induce autophagy, ...turnover of cytosol and organelles in yeast. Autophagy,4(2), 205-213. doi:10.4161/auto.5331. PubMed PMID: 18094608...PMID: 20144757. PubMed Central PMCID: PMC2852113. 4. Klionsky, D. J., Cuervo, A. M., & Seglen, P. O. (...information about the contents of the autophagosome nor does it provide information about what is being...higher pH of the vacuole. Read on to learn more about prior methods for studying autophagy and how Rosella... (as opposed to giving more fine-grained detail about what proteins or organelles are being degraded) ...autophagy, but, like the protein degradation assay above, it’s slow and only measures bulk autophagy. Morphological...

...alpha L [IPI-TS2/4] Integrin alpha-L Human IgG1 Rabbit Anti-Integrin alpha L [IPI-M17/4] Integrin alpha-L...alpha L [IPI-TS2/4] Integrin alpha-L Human IgG2a Mouse Anti-Integrin alpha L [IPI-M17/4] Integrin alpha-L...roles of individual integrins. The Ha lab — in collaboration with the Springer lab, the Jaumouillé lab, the...recombinant antibodies and plasmids. To encourage collaboration and education in protein science, IPI hosts ...events and training courses on technologies and laboratory techniques related to antibody discovery. You...the Institute for Protein Innovation. She writes about all things IPI, from the scientists and scientific...

... Fishell 83897 pAAV-hDlx-GqDREADD-dTomato-Fishell-4 hDlx Activator DREADD Fishell 98747 pAAV-FLEX-EGFPL10a...serotype from the Alla Karpova and David Schaffer laboratories. These viral vector preparations were produced...Janelia Research Campus viral project team in collaboration with David Schaffer (University of California...

...component plasmids on the Addgene website. Step 4: Naming your assembled plasmid and generating the ...the vector is to your liking, your last step, Step 4, is to give the assembled plasmid a name and generate...assessment of emerging CRISPR technologies. Cell Genomics, 4(3), 100519. https://doi.org/10.1016/j.xgen.2024.100519...Destination vector (dropdown options depending on above selections) Module set (the set of components you...vectors) Target type (drop down options depending on above selections.) CRISPR enzyme (Cas9 [sp], Cas9 [other...

...assembles the final targeting vector (Fig. 1, Step 4), and co-injection of the targeting vector and a Cas9...Modification in Caenorhabditis elegans.” Genetics. 202(4) (2016):1277-88. PubMed PMID: 26837755. PubMed Central...modular base-specific gene tagging using a ligase-4-dependent mechanism.” Nat Commun. 7 (2016):12338. ...tagged gene. Most insertions can only be detected by labor-intensive processes like PCR or by evaluating visual...repair after a CRISPR/Cas9 induced break. Learn more about these constructs and the cell lines they’ve been...Additional Resources on the Addgene Blog Learn about Fluorescent Protein Timers Check out Photoactivatable...

... ShadowR, a novel FLIM-FRET acceptor Figure 4: ShadowR is used as a FRET acceptor to mRuby2 or mScarlet...energy transfer (FLIM-FRET) in living cells (Figure 4). They created ShadowR by modifying the surface of...Reports, 9(1). https://doi.org/10.1038/s41598-019-48604-4 Salih A, Larkum A, Cox G, Kühl M, Hoegh-Guldberg O... Fluorescent Protein 101 blog series Learn more about FRET Find protocols and tips for making plasmid ...

...research more inclusive. CBE Life Sciences Education. 13(4):602-606. Brian, D. 1996, Einstein: A Life. Chapter...attitudes in CUREs. CBE Life Sciences Education. 21(4):ar74. Duboue, E., Kowalko, J., Keene, A. et al. 2022...and neurobiology. CBE Life Sciences Education. 15(4):ar55. Ma, Y., Mukherjee, S., Uzzi, B. 2020. Mentorship...in a year-long biochemistry laboratory : Part II – research-based laboratory – a semester-long research...are often associated with rote memorization of metabolic pathways (precisely the type of exercise Einstein...had the opportunity to talk with Provost recently about his development of what is now commonly termed a...into one of my own courses, I was excited to hear about how he developed his program. Initially, Provost...