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We narrowed to 1,018 results for: REN

Showing: 441 - 480 of 1018 results

  1. CRISPR Challenges: Standardization and Homology Directed Repair

    Type
    Blog Post
    ...work demonstrates that different repair template strategies activate different HDR sub-pathways like synthesis-dependent...live updates! My most popular tweet from the conference is shown above. The blog post I mentioned, John...post-CRISPR cleavage When I first looked at the conference agenda, I was surprised to see that it would ...Having had time to digest my notes, since the conference, I think I may have finally grasped just how ...Subscribe to CRISPR updates on our blog to keep current with the latest and greatest! Addgene resources...

  2. Why Add Sucrose? Improved Yields for Adeno-associated Virus Preparation

    Type
    Blog Post
    ...redesigning viral packaging plasmids, investigating different viral production strategies or developing novel...silver stain approaches we show that there is no difference in purity or infectivity between the untreated...GFP-expressing AAV particles we did not observe any difference between untreated and sucrose treated cells. ...it will be exciting to test this approach with different capsids. Most importantly, because our AAV yields...spending less time and resources resupplying our current inventory and more time providing new materials...materials to the research community. References 1. Coroadinha, A. S., et al. "Effect of osmotic pressure on...

  3. Interview: Ed Boyden on Optogenetics, Neuroscience, and the Future of Neuroengineering

    Type
    Blog Post
    ...specific problem. Every species is so different – an awake monkey is different from a cell in a dish. I always...want to activate two sets of neurons and see how different parts of the brain or the cells within them work...Boyden: I think because neuroscience has so many different goals: to understand how a mouse changes emotions...find ways to make these tools more selective for different ions. Right now, we can modulate electricity. ... click the link in the Addgene Alerts box. References: Independent Optical Excitation of Distinct Neural...

  4. Sequencing Options for CRISPR Genotyping

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    Blog Post
    ...This post was contributed by guest blogger Søren Hough, a Biochemistry PhD candidate at the University...double-stranded breaks (Tsai et al. 2015). Additionally, current algorithms do not take into account other elements...sequencing (Hendel et al. 2015), a significant difference as CRISPR experiments move closer to the clinic...and Joung 2016. Many thanks to our guest blogger Søren Hough. We additionally thank Monica Sentmanat, Victor...Ajetunmobi for their contributions to this work. Søren Hough is a Biochemistry PhD candidate in the laboratory...https://magazine.scienceforthepeople.org/.     References 1. Fu, Yanfang, et al. "High-frequency off-target..."Quantifying on-and off-target genome editing." Trends in biotechnology 33.2 (2015): 132-140. PubMed PMID...

  5. Predicting Adverse Reactions to Monoclonal Antibody Drugs

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    Blog Post
    ... and current Scientist, DNA Sequencing and QC at Addgene.  References and resources References Mosch... T-cells or monocytes. PBMCs can be further differentiated into dentritic cells or macrophages through... 3-D models can be applied to corroborate the occurrence and extent of the adverse drug reaction. This...of Arbeitsgemeinschaft fur Pharmazeutische Verfahrenstechnik e.V, 130, 128–142. https://doi.org/10.1016...

  6. Viral Vectors 101: Inverted Terminal Repeats

    Type
    Blog Post
    ... interest, and a polyA signal. Created with biorender.com. ITR structure  ITRs are symmetrical 145-nucleotide...resolution site (trs) are noted. Created with biorender.com. ITR modifications and types  Since ITRs are...expression can be altered by using ITR sequences from different AAV serotypes that vary in their RBE and T-shaped...multiple AAV serotypes (Wilmott et al., 2019), and different combinations of ITR and capsid serotypes vary ...restriction digest methods.    Resources and references More resources on the Addgene blog Viral Vectors...Addgene.org Addgene's AVV guide AAV Viral Preps References  Earley, L. F., Conatser, L. M., Lue, V. M., ...

  7. Quantifying DNA? Here are Five DNA Quantification Methods to Consider

    Type
    Blog Post
    ...the liquid. Molecules absorb different wavelengths of light to different degrees and many molecules have...spectrophotometer measures these absorbances using UV-transparent cuvettes. First you measure the absorbance of...the ladder. You then run samples of your DNA at different dilutions and quantify based on band intensities...elute and store your plasmid DNA Learn about the different grades of plasmid DNA preps Resources on Addgene.org...Addgene.org Visit the Molecular Biology Reference Find plasmids for your research Browse protocols for ...

  8. Science Career Options

    Type
    Blog Post
    ...are normally available (as sectors such as forensics currently have too many applicants compared to the ...Michael Koeris' Journey from Grad Student to Entrepreneur Science careers divided by the most pertinent...types of skills show that you fully explore the different options available and then decide on a course ...full time) and what life is like for someone who currently in the position. Questions you’ll want to find...Michael Koeris' Journey from Grad Student to Entrepreneur Learn to Network and Make the Contacts for Your...

  9. Improving Transduction Efficiency with Exosome AAVs

    Type
    Blog Post
    ...gradients. The current purification protocols for isolating exo-AAVs rely on differential centrifugation...saw that these AAV particles come in a slightly different shape. These AAV particles are often found with...able to infect more cells (as demonstrated in different cellular environments such as retina or glioblastoma...Exo-AAVs have improved transduction profiles in different AAV serotypes both in vitro and in vivo when compared...to inactivation through anti-AAV antibodies. References György, Bence, et al. "Naturally enveloped AAV...

  10. Quick Guide to Working with Drosophila Part 2: Controlling Gene Expression in Flies with Gal4/UAS

    Type
    Blog Post
    ... Gal4 lines and study gene expression in many different tissue types! Although putting a gene of interest... overexpressed in several cell types by using different Gal4 drivers. It is much easier to mate flies ...one gene in one cell type and another gene in a different cell type (6–8). Although it’s often very easy...the meantime, be sure to browse the publication references and web links from my first post and start designing...jonchowphd but he still reads “#” as a “pound symbol”. References 1. A. H. Brand, N. Perrimon, Targeted gene expression...systems in Drosophila: A synthesis of time and space, Trends Genet. 20, 384–391 (2004). PubMed PMID: 15262411...

  11. DAP arrays for multiplex gene editing

    Type
    Blog Post
    ...either from human or plant sources) in both the strength as a promoter and the activity to release gRNAs...heart disease, and type 2 diabetes, across four different types of human cells, including T cells and hematopoietic... interest with only minimal genetic payloads. Different from existing multiplex array architectures, including...when repetitive tRNA sequences must be avoided, different tRNAs with optimized 5’ leader sequences can be... lab of Xue Sherry Gao at Rice University.    References and Resources More resources on the Addgene blog...Adding Precision and Flexibility to CRISPR Editing References Yuan Q, Gao X (2022) Multiplex base- and prime-editing...

  12. R Bodies: Membrane-Rupturing Microscopic Tools

    Type
    Blog Post
    ...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...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...

  13. Components of CRISPR/Cas9

    Type
    Blog Post
    ... 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...sequence.   Now that you know about all the different components of CRISPR, It's time to start thinking...

  14. Resume Writing for Non-academic Science Careers

    Type
    Blog Post
    ... is it different from a CV? Get familiar with the general format of a resume and the different presentation...If the job posting says “French-speaking is an asset,” mention “speaks French.” Employers usually don'...you volunteer to do science outreach and teach children about science? Or perhaps you have presented in... science outreach and writing was how I got my current job as a Communications Coordinator. Wondering ...

  16. Sweating the Small Stuff: Details in the Lab

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    Blog Post
    ...the answer isn't to work harder - it's to work differently.  Tools to Try  Checklists Found in surgical ...and your workflow can have a huge impact. Test different approaches, like whether it's better for you to...as a Do Not Disturb sign, even if the headphones aren’t actually playing music. Nontraditional cues are... more than anything else I tried, made a huge difference in my ability to focus, especially on days when...what can be a rather vicious cycle.  Everybody’s Different   The truth is, there is no One True Way to Error-Free...the detail-driven daily grind of science.      References and Resources Images used Roberson Lab, UPenn...

  17. Fluorescent CRISPR Reporters: SRIRACCHA and GEmCherry2

    Type
    Blog Post
    ...These cells are then transfected with several different constructs for the reporter to work and to make... visual read out allows for easy screening of different gRNAs by simply changing the gRNA used when transfecting...labor-intensive, require multiple fluorescent proteins, and aren’t that efficient. To overcome these issues the Denham...concept the Denham lab compared editing at three different genomic sites in the AAVS1 locus using GEmCherry2...GEmCherry2. In this experiment they saw a clear difference in mCherry expression using FACs sorting, confirming...mutant of interest, give these methods a try.  References Højland Knudsen C, Ásgrímsdóttir ES, Rahimi...

  18. Enabling Precision Functional Genomics with the Target Accelerator Plasmid Collection

    Type
    Blog Post
    ...approach. One central bottleneck is that most recurrently mutated cancer genes are rare and most of the...found in genome sequencing studies (as well as reference cDNA clones mapping to well-annotated cellular...signaling pathways as part of our Genetic Pathway Reference Set. These Gateway-compatible entry clones are...cancer mutant collection and the genetic pathway reference collections are described in this paper Systematic...Central PMCID: PMC5386591.  The genetic pathway reference collection is described in this publication ...Comprehensive Set of MAPK1/ERK2 Missense Mutants. Brenan L et al. Cell Reports, 2016. PubMed PMID: 27760319...

  19. Hot Plasmids - June 2019 - Optogenetics, Acoustic Reporter Genes, microRNAs, and the CRISPR-Cas9 system CHIME

    Type
    Blog Post
    ...help define the different cells present in a mixed population, or identify differentiation and disease states...essential for synaptic transmission. Cleavage of VAMP2 renders it inactive, thus inhibiting neurotransmitter release...some genes can change depending on the location. Current methods for visualizing the location and function...challenges such as the inhibition of immune cell differentiation. Furthermore, TALEN and ZFN require custom ... any immune lineage without affecting cell differentiation in a high-throughput manner. They designed ...

  20. Will You Be My Mentor? Finding and Asking for Mentoring Support

    Type
    Blog Post
    ...help us in different ways and in diverse areas. It is helpful to have mentors from your current organization...that you are an expert networker (or if you just aren’t doing it at all yet) watch my webinar, “Not Networking...you identify specific skills you need for your current position Understand your interpersonal challenges...someone you trust. A good coach can make a big difference, if you can find one. Create your own peer mentoring...with progress. Tell them how their advice made a difference, helped you make a change for the good or resulted...

  21. Special Delivery: Fluorophore Targeting for FRET Studies

    Type
    Blog Post
    ...targeting the donor and acceptor fluorophores to the different biomolecules you’d like to test for interaction...that the targeting specificity requirements are different for the donor and acceptor fluorophores. The donor...acceptors are delivered to the wrong address in a different city, you can still develop a successful FRET-...signal is unaffected  Due to these fundamental differences in targeting specificity for donor and acceptor...blogger James D. Fessenden! James D. Fessenden is currently an Assistant Professor in the Anesthesia Research...channels involved in skeletal muscle contraction.   References 1. Degorce, François, et al. "HTRF: a technology... theoretical aspects and recent applications." Current Chemical Genomics and Translational Medicine 3.1...

  22. Antibodies 101: Fab Fragments

    Type
    Blog Post
    ... for your specific scenario.    References and resources References Bates A, Power CA (2019) David vs...Constant domain. H = Heavy chain. Created with biorender.com     The modularity of antibodies means ...is relatively easy to physically divide up the different regions, both through genetic and proteolytic ...broken up, there are a few commonly available different fragments that retain antigen binding (Figure ...common antigen binding fragments. Created with biorender.com.   Why Fabs?  Antibody fragments have a ...

  23. Antibodies 101: Conjugation

    Type
    Blog Post
    ...good base from which to dive deeper!References and resources References Bendall SC, Nolan GP, Roederer M,...love FPbase’s Spectra Viewer tool for comparing different fluorophores. We also touched on fluorophores ...chemical coupling can be achieved through several different mechanisms. Most take advantage of chemical groups...interactions with other peptides. Created with BioRender.com.     Many biologists can lead perfectly ...PK (2012) A deep profiler’s guide to cytometry. Trends Immunol 33:323–332. https://doi.org/10.1016/j.it...

  24. All in a Twist: dsRNA

    Type
    Blog Post
    ... disease, and much more! Resources and References References Wang, I. X., Grunseich, C., Fox, J., et ...replication process within the host. dsRNA viruses are inherently double stranded, while ssRNA viruses can generate... dsDNA viruses yield dsRNA through a slightly different mechanism thought to be associated with bidirectional...transcription, RNA is synthesized from a DNA template, inherently generating RNA:DNA intermediates in the process...blog CRISPR 101: RNA Editing with Cas13  RNA Interference in Plant Biology  ...

  25. CRISPR 101: Cas9 vs. The Other Cas(s)

    Type
    Blog Post
    ... put your experiments Nsync.   References and resources   References Dolan, E., Wang, Hou, Zhonggang,...while also highlighting their similarities and differences to Cas9. DNA targeting Cas enzymes Cas9 – The...Originally classified as Cpf1, Cas12’s primary difference from Cas9 is that it generates staggered breaks...(RNA knockdown, RNA editing, etc.). The main differences between Cas7-11 and Cas13 are 1). non-specific... Cas While Cas9 may be the most adaptable Cas currently available, the other Cas proteins should not be...

  26. The Breast Cancer Microenvironment: A Tumor’s Backstage Team

    Type
    Blog Post
    ...progression, enzymes and metabolism reprogramming aren’t the whole story. It’s essential for a tumor to ... integrins (cell-adhesion proteins) targeting different organs. This means that exosomes not only promote...microenvironment’s malignancy but also incite different organs to become malignant. Exosomes also contain...guest blogger, Bárbara Pinho! Bárbara Pinho is currently a science communicator at the Portuguese science...issues. Follow her on Twitter @BrbaraPinho6.      References Allinen, Minna, et al. "Molecular characterization...stromal caveolin-1 expression predicts early tumor recurrence and poor clinical outcome in human breast cancers...

  27. dTAG - You're it!

    Type
    Blog Post
    ...ortho-AP1867) via various chemical linkers. Our current lead dTAG molecule is dTAG-13 due its immediate...selectivity, broad utility for degrading targets in different cellular compartments, and in vivo utility. dTAG...dTAG molecule toxicity in new cell lines Treat parental cells (or cells expressing a control -- for example...expression of these cereblon neo-substrates varies in different contexts (for example, IZKF1 and IZKF3 are cell-type...technology. Follow him on twitter @behnamnabet.   References Nabet et al. The dTAG system for immediate and... 14(5): 431-441. PubMed PMID: 29581585. This reference contains the full description of the dTAG system...

  28. It’s Bioelectric! An Exciting Interdisciplinary Field of Research

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    Blog Post
    ...override genetically encoded patterning. Based upon trends in studies over the past several years, it seems...negative) the action potential of a cell, the more differentiated and quiescent it is, and the more depolarized... toggled between open and closed states using different wavelength light pulses. Other resources from...bioelectricity, bioelectric researchers in many different fields came together to found the journal Bioelectricity...regeneration, medicine, and synthetic bioengineering. References Chernet, Brook T., et al. "Use of genetically...for regenerative and synthetic bioengineering." Current opinion in biotechnology 52 (2018): 134-144. PubMed...

  29. Gendered Innovations: Why Does Sex of the Cell Matter?

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    Blog Post
    ...for men. Not only does developing a drug in the current market cost billions—but when drugs fail, they ... relevant? Research shows that there are sex differences in the therapeutic capacity of stem cells. Yet...male stem cells. Nor did they detect important differences in function between male and female cells. The...in basic research.  In addition to the studies referenced above, another excellent resource for research...population. To learn a little more about where differences in effects on the sexes come from, one clue or...expert on gender in science and technology and currently directs the EU/US Gendered Innovations in Science...

  30. CRISPR Meets Synthetic Biology: A Conversation with MIT’s Christopher Voigt

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    Blog Post
    ... to what’s happening at different points within the wood. There are different types of cellulose fibers...number of DNA binding proteins with Cas9 guided by different promoters. The challenge in building genetic circuits...then you’d need the ability to turn on genes at different times. It requires more complex synthetic networks...circuits, they are extremely orthogonal. It’s a different perspective and you can layer them for more complicated...for multi-input CRISPR/Cas9 genetic circuits References: Nielsen AA, Voigt CA. Multi-input CRISPR/Cas...

  31. Viral Vectors 101: Chemogenetics

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    Blog Post
    ... and some, such as varenicline, are already approved for clinical use. Varenicline has been further modified...updated by Jason Nasse in July 2023.  References and Resources References Chen X, Choo H, Huang X-P, Yang X...to consider that the same receptor can elicit different responses depending on what cell type it is expressed... signaling. The two main classes of receptors currently used in chemogenetics are DREADDs and PSAMs. ...

  32. Plasmids for Endogenous Gene Tagging in Human Cells

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    Blog Post
    ...portal. The plasmids should be functional in many different human cell types, and while most of the genes ...protein from the untagged allele. Created with BioRender.com. The first step in creating the plasmids...GFP tag sequence to be inserted. Created with BioRender.com. A key feature of our methodology is the...enable us to effectively utilize the host cell’s inherent homology directed repair (HDR) process following...break and introducing the tag. Created with BioRender.com. Plasmids, plasmids everywhere To find out...Allen Institute website at alleninstitute.org. References 1. Roberts, Brock, et al. "Systematic gene tagging...

  33. Adenoviral Vector Production and Troubleshooting

    Type
    Blog Post
    ...for production of Gutless AdV can be found in reference 3. Production, amplification, and quality control...please see Addgene’s Adenoviral Guide webpage, and references 1, 2.*Pro-Tip*: once the correct recombinant ...specific for a viral DNA sequence. *Pro-Tip*: Ad5 reference material should be used as an internal control..., high-titer rAdV in your experiment. Enjoy! References 1. Luo, Jinyong, et al. "A protocol for rapid...practical guide for using the AdEasy System. 3. Jager, Lorenz, et al. "A rapid protocol for construction and ...  Karen Guerin...

  34. Rabies and Neuronal Tracing

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    Blog Post
    ...forms of these viruses typically used in research aren’t able to replicate, and for a virus to cross a synapse...Watabe-Uchida et al., 2012). They could also quantify the strength of the inputs by counting the monosynaptic inputs...more inputs than was previously thought. The differences in the inputs to each of the two populations ...populations could be used to explain the differences in the functions of these two neuronal populations. Overall...Overall, this study laid the foundation for how different anatomical regions of the brain receive inputs and...complex questions of how the brain operates. References 1. Callaway EM. Transneuronal Circuit Tracing...

  35. Hot Plasmids - October 2020

    Type
    Blog Post
    ...number, generally very small, and can contain different neurotransmitters. Using genetically encoded fluorescent... brain.  Figure 1:Schematic diagram of different FingR constructs and experimental results. Image... of the cell. By combining vectors expressing different fluorescent proteins targeting inhibitory or excitatory...The team validated this tool by targeting six different genes whose protein products have distinct localizations...Figure 2: Split-wrmScarlet labeling proteins with different subcellular locations. Image from Goudeau et al...consistently predicted the cell cycle speed across these different cell types. In one example, H2B-FT was expressed...

  36. Story of a SynBio Startup: RevBio's Epiphany (or Lack Thereof)

    Type
    Blog Post
    ...order to head to Ireland and take a chance as entrepreneurs. To make things more interesting, we are working...academic labs. Being unable to publish or attend conferences makes it very challenging to compete in today...pressure to publish and more freedom to attend conferences, but after receiving her degree she was faced...ideas about things we could do for work, and they weren’t restricted to science jobs. Keira’s an optimist...career, but it was enough to start us down the entrepreneurial path. We threw some of our own money at this...thinking about making the switch from academia to entrepreneurship, share your thoughts with us in the comments...

  37. Viral Vectors 101: Producing Your rAAV

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    Blog Post
    ...updated by Ina Ersing in August 2023 References and resources  References  Aurnhammer, C., Haase, M., Muether...required adenoviral helper genes are supplied on two different packaging plasmids: the pHelper plasmid containing...titering assay, you’ll need to understand how different titers are measured. A TCID50 measurement reports...have a high-quality plasmid standard and rAAV reference material of a known titer on every qPCR run. Note...of a recombinant adeno-associated virus type 2 Reference Standard Material. Human Gene Therapy, 21(10),...

  38. Viral Vectors 101: Voltage Indicators

    Type
    Blog Post
    ...voltage indicators reviewed.   References and Resources   References Beck C, Zhang D, Gong Y. Enhanced... These types of sensors can have a variety of different components, but all contain a FRET pair, a genetic...specifically localize to the membrane, otherwise they aren’t all that useful. The cellular localization component...target – and this serotype can be selected to preferentially target your cell type of interest. Viral pseudotyping...powerful tool that swaps capsid proteins from different viruses to optimize them for targeting your cell...

  39. Multiplexed Capture of Promoter-enhancer 3D Chromatin Structures Using CRISPR

    Type
    Blog Post
    ...sequencing data. While “pictures” generated with current 3C methods provide useful information about chromatin...compare results across multiple targets because different CAPTURE cell lines will have varying levels of...lines Cell lines, primary cells Table 1: Key differences between original CAPTURE and CAPTURE 2.0 methods...that occur during development and cellular differentiation. Using a cell line model, the Xu Lab tracked...enhancer-promoter interactions during erythroid differentiation. CAPTURE 2.0 captures chromatin's dynamic role....0 re-writes the script on gene regulation. References Han, J., Zhang, Z., & Wang, K. (2018). 3C and...

  40. Addgene's Cancer Collection Pages Connect the Research Community

    Type
    Blog Post
    ...cellular pathways involved in the development of different cancers and identifying the roles of key proteins...approaching the Ras problem by exploiting the different mutant versions of Ras that are most prevalent...function of any of these genes through several different applications such as fluorescent protein fusion... study it (as seen in the screenshot below). Currently, the page includes interactive maps for the Ras...interests in education and scientific writing.     References 1. Martz, Colin A., et al. "Systematic identification... PMID: 24651010. 3. Ledford, Heidi. "The ras renaissance." Nature 520.7547 (2015): 278-280. PubMed PMID...
Showing: 441 - 480 of 1018 results