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We narrowed to 1,006 results for: RING

Showing: 481 - 520 of 1006 results

  1. Networking Follow-up: I Met a Person. Now What?

    Type
    Blog Post
    ...organization, mentoring group, or department. Even if it doesn’t work out, it’s flattering to ask.  Show...Relationships need nurturing. Perhaps you’ve met someone interesting at an event. Or, perhaps you have...There is a person on LinkedIn I once helped by answering one simple question so I accepted the connection...

  2. Expanding the Targeting Scope and Editing Efficiency of Adenine Base Editors

    Type
    Blog Post
    ... adenine base editor Lower-stringency PANCE, followed by higher stringency PACE, resulted in the evolution...which results in less overall dilution and less stringent phage selection. Figure 2: Phage-assisted...base editors? Targeted evolution of naturally occurring enzymes and functional domains speed up the development...

  3. Antibodies 101: Isotypes

    Type
    Blog Post
    ...immune response. There are three main pathogen clearing functions or effector functions of the constant...each of the isotypes. IgM This isotype is produced during a primary immune response because all B cells begin...found in tissues. IgG is the predominant isotype during a secondary immune response, which is when the ...constant domains. This switching helps B cells adapt during the course of an immune response since it also ...

  4. 15 Years of Addgene: The Top 15 Plasmids

    Type
    Blog Post
    ...for supporting scientific sharing. We’re very excited for more reagent sharing projects in the years to...15 years of plasmid sharing has certainly taken us on many adventures...From moving office locations ...Cas9 (SpCas9) has been widely popular in genome engineering. For use in human cell lines, the Zhang lab codon-optimized...

  5. Viral Vectors 101: Viral Vector Elements

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    Blog Post
    ...S.H., Varmus H.E. Retroviruses. Cold Spring Harbor (NY): Cold Spring Harbor Laboratory Press; 1997. Principles...ITRs are perfectly symmetrical to allow for this pairing to occur. Like LTRs, the ITRs define the limit ... supplied by the production cell line (HEK-293) during production. Fun fact: There are 57 human adenovirus...

  6. Viral Vectors 101: Biosensors

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    Blog Post
    ...expanded for multiplexed imaging for advanced monitoring of different factors, in different or the same...stimuli and as well as changes in cellular dynamics during so-called housekeeping activities.     ...cell (sub)types for expression and downstream monitoring. Nervous about making a virus for your biosensor...Biosensors for Neurotransmission and Neuromodulation: Engineering and Applications. Frontiers in Cellular Neuroscience...

  7. Developing a cold tolerant E. coli using specialized chaperones

    Type
    Blog Post
    ...milk processing, fruit juice clarification, meat tendering, and oligosaccharide synthesis. A major incentive...insoluble granules. We overcame this problem by engineering a plasmid which contained cold tolerant chaperones...editing for psychrophilic systems as well as engineering and regulating psychrophilic recombinant proteins...synthetic biology experiment Read our Genome Engineering Guide ...

  8. CRISPR 101: Non-Homologous End Joining

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    Blog Post
    ...advantage to using the CRISPR/Cas system for genome engineering is the fact that Cas9 can be easily programmed...initial cut, the next steps in the process involve repairing chromosomal DSBs. It is important to know that...then ligates compatible DNA ends together, thus repairing the break. This is a simplified, streamlined ... ligation step determines how ends are resolved during nonhomologous end joining. Nat Commun 5: . https...

  9. Pooled CRISPR Libraries Offer Genome-Wide Control for Large-Scale Functional Screens

    Type
    Blog Post
    ...rely on genetic engineering to optimize humans for CRISPRi and CRISPRa.” Before considering the use of pooled...that they could be used by people with little bioengineering expertise? “This is the next step in high-throughput...activating any human gene, go to http://sam.genome-engineering.org/. Crystal structure of Cas9 in complex with...

  10. Deep Dive: qPCR

    Type
    Blog Post
    ...to double-stranded, and finally, drop once more during a final temperature increase. If there is a single...should only be one peak, as everything should be denaturing or annealing at the same temperatures. If there.... Flipping your experimental and control values during analysis can allow you to see this fold change ... vol. 1160, DOI 10.1007/978-1-4939-0733-5_3, © Springer Science+Business Media New York 2014Bustin S; ... as Reference for qPCR Gene Expression Analysis During iPS Reprogramming Process. Sci Rep 8, 8716 (2018...

  11. Hot Plasmids - March 2020 - base editors, GEVI, MoClo, and optogenetics

    Type
    Blog Post
    ...editors at Addgene. Levy et al., Nature Biomedical Engineering, 2020. PubMed PMID: 31937940. Detection of neural... prosthetic group PCB, PhyB and PIF6 dimerize, bringing each terminus of Cre together to restore Cre activity...to red light induces PhyBΔ and PIF6APB binding, bringing the two halves of Cre together. Image from Eisenhoffer...as well as in different stages of development, ensuring the breadth and versatility of this tool. Find...

  12. What's New in CRISPR - November 2021

    Type
    Blog Post
    ... system. The CasMINI system was developed by engineering the Cas12f scaffold gRNA and making strategic...the gRNA scaffold through five rounds of gRNA engineering.  They did not modify the Cas12f1 enzyme. The...cargo RNA, and fusogen in a modular system for delivering gene editing tools into cells. Image adapted ... within the FAH gene was precisely corrected, restoring expression in the liver.  Read the paper in Nature...

  13. Viral Vectors 101: An Introduction to AAV

    Type
    Blog Post
    ...vector into the culture medium during adeno-associated virus manufacturing." Human gene therapy 21.10 (2010...researchers speak about AAVs, they are actually referring to rAAVs. ITRs and serotypes The names of AAVs...different ages." Retinal Degenerative Diseases. Springer International Publishing, 2016. 501-507. PubMed...

  14. Rosella: A Fluorescent pH-Biosensor for Studying Autophagy

    Type
    Blog Post
    ... Mark Prescott. This system was developed for monitoring and analyzing autophagy of cytosol and organelles.... Autophagy is then induced and quantified by measuring the amount of radioactivity in the culture supernatant...autophagy markers. It’s particularly useful for comparing bulk autophagy vs. mitophagy, the targeted autophagy...Cuervo, A. M., & Seglen, P. O. (2007). Methods for Monitoring Autophagy from Yeast to Human. Autophagy,3(3)...

  15. CRISPRainbow and Genome Visualization

    Type
    Blog Post
    ...with the previously established physical map. In comparing pairs of targets ~2 Mbp apart, they noticed that... magenta, depending on the hairpin combination bringing the total number of individual colors to 6. A ...Addgene. 4. Kiani, Samira, et al. “Cas9 gRNA engineering for genome editing, activation and repression... Mary Gearing...

  16. All in a Twist: dsRNA

    Type
    Blog Post
    ...are typically contained within the mitochondria, barring a loss of mitochondrial integrity that would release...stranded structure produced is known as an R-loop. During transcription, RNA is synthesized from a DNA template...transcription of thousands of genes is constantly occurring throughout a single cell, so these nucleic acid...DNA hybrids Persistent RNA:DNA hybrids that arise during transcription are typically recognized by RNase...

  17. Enhancing CRISPR Targeting Specificity with eSpCas9, SpCas9-HF1, & HypaCas9

    Type
    Blog Post
    ...effects” have included using a Cas9 nickase variant, lowering Cas9 expression, and truncating gRNA sequences...decrease the off target effects of CRISPR/Cas9 by altering the nuclease activity of the Cas9 nuclease itself...excited to see continued improvements in genome engineering! References 1. Slaymaker, Ian M., et al. "Rationally...Plasmids Browse our CRISPR Guide Pages Find Genome Engineering Resources ...

  18. New Tool for Lineage Tracing: The ClonTracer Library

    Type
    Blog Post
    ... populations resistant to drug treatments arise during treatment or are preexisting in the parent population... drug treatment and that these are selected for during the treatment process. The authors went on to show... “I spend a lot of time packing, shipping, and sharing protocols one by one,” and she is excited to deposit...Addgene because it is a “really great way of broadly sharing these kinds of reagents.” Here at Addgene, we’re...

  19. Career Coaching for Scientists: Why and Where Do I Find One?

    Type
    Blog Post
    ...infrastructure are not always as effective in mentoring for this transition. Second, a coach will help...through on the actions you committed to. Finally, hiring a paid coach will make you take the experience ...Joanne's Reddit AMA I am a strong advocate for peer mentoring, but volunteer mentors are only able to go so ...areas of specialty from leadership development to mentoring for women and under-represented minorities. Her...

  20. Meet an Addgenie: Insights into Grad School, Career Paths, and More

    Type
    Blog Post
    ...and a Spanish minor, it’s time for me to start considering my future path. Talking to Addgenies has helped...it solidified some concepts that are heard often during undergrad. Lauren Flynn is the Addgenie behind... publish their research and are not so keen on sharing their results in fear of getting scooped, she said...her focus is finance, it is just as important to bring love and fun to the office. Main takeaways from ...

  21. Plasmid-based Recombinant Monoclonal Antibodies: What They Are and Why You Should Be Excited About Them

    Type
    Blog Post
    ...still have a shelf full of CDs that have been gathering dust since the digital music revolution hit. I...not shared. For instance, through open plasmid sharing, the CRISPR community has created hundreds of useful...Antibodies: a Laboratory Manual, 2nd Edition. 2104. Cold Spring Harbor Laboratory Publications, New York. Helma...Monoclonal Antibodies for Clinical Use. In: Antibody Engineering. InTech Additional resources on the Addgene blog...

  22. Popular Retroviral Vectors and Their Uses in Scientific Research

    Type
    Blog Post
    ...Therefore they make excellent tools for genetic engineering. There are several different viral vectors to... and Adeno-associated virus (AAV). If you are wondering which virus fits you experiments best have a look...retroviral vectors can regulate gene expression by delivering shRNA or miRNA. The ever expanding CRISPR field...our CRISPR reagents. Please check www.genome-engineering.org for the most up-to-date information. Check ...

  23. CRISPR in the Clinic

    Type
    Blog Post
    ...involve editing healthy donor-derived cells and delivering them to the patient.   Current CRISPR Therapies... preliminary results (Dimitri et al). CRISPR engineering of CAR-T cells is particularly promising because...Early-stage trials showed a high success rate in clearing misfolded protein levels as measured in the blood...2013). Dimitri, A., Herbst, F. & Fraietta, J.A. Engineering the next-generation of CAR T-cells with CRISPR-Cas9...

  24. Fluorescent Proteins 101: Introduction to FRET

    Type
    Blog Post
    ...Quantification of Förster resonance energy transfer by monitoring sensitized emission in living plant cells." (... for studying protein-protein interactions by measuring the change in FRET efficiency, from which conclusions...cascades, quantify mechanical tension (a molecular “spring”) (Meng et al., 2008), and monitor neuronal activity...Sustarsic and Kapanidis 2015), which may lead to monitoring of these processes in live animals (Hirata and...

  25. Plasmids 101: Modular Cloning

    Type
    Blog Post
    ...Most things in biology are not so simple, and engineering complex cellular behaviors requires making complex...output, etc., which can support a larger cellular engineering goal, several of which are described below. Multigene...assembling an array of targeting constructs and delivering them to these organisms via another optimized... an array of distinct goals within metabolic engineering, synthetic product production, and gene editing...

  26. Adapting Toehold Switches to Detect Zika Virus

    Type
    Blog Post
    ...risk for the project by initially designing and ordering several dozen versions of the toehold switches...successful sensors to draw on as our assays grew more stringent, and it avoided any days of lost productivity ... virus genome with Addgene to facilitate rapid sharing of these tools with the Zika virus research community...product development, the regulatory process, and manufacturing so that we can get the diagnostics into the ...

  27. Cpf1 Update: Comparison to Cas9 and NgAgo

    Type
    Blog Post
    ...shortcomings. Cpf1’s preferred PAM is 5’-TTN, differing from that of Cas9 (3’-NGG) in both distribution...clinical applications require HDR over NHEJ, so comparing HDR efficiency across CRISPR nucleases and NgAgo...insertion mediated by Cpf1-generated overhangs. Exploring the efficiency of these two processes will help... Mary Gearing...

  28. Reaching out to China: Canton Nucleic Acids Forum (CNAF) 2015

    Type
    Blog Post
    ...lab is also exploring the use of cas9 proteins from different species for genome engineering. The final...will lead to advances in human health. It was inspiring to hear how the genome project has already had...complicated conformation “not like DNA which has a boring structure.” That certainly summed up this conference—nucleic...

  29. Experimenting in an Open Source lab: from CRISPR to Cats

    Type
    Blog Post
    ...function as centres for peer learning and knowledge sharing, in the form of workshops, presentations, and lectures...looking at how to re-use it as a platform for monitoring homemade microfluidic chips using the fantastic...members and the public cheap access to personal manufacturing tools – a 3D printer, a laser cutter, woodworking... and an instructor at Biocurious.org, Derek is bringing cutting edge technologies to Victoria, BC. Follow...

  30. Adenoviral Delivery of CRISPR/Cas9 Aims to Expand Genome Editing to Primary Cells

    Type
    Blog Post
    ... Manuel Gonçalves; made with BioRender.   Delivering gRNA and Cas9 using adenoviral vectors As compared...published in Nature Methods the same year that delivering RNA-guided nucleases or TALENs together with ...the lab of Andrea Ventura at the Memorial Sloan Kettering Cancer Center did to create a model of Eml4–Alk-driven...Stanchina E, Lowe SW, Ventura A (2014) In vivo engineering of oncogenic chromosomal rearrangements with ...

  31. The Scientific Conference Poster Session: Tips for Success

    Type
    Blog Post
    ...re a presentation veteran, but have difficulty answering questions. Or perhaps you’re simply attending,...snacks to keep you energized throughout the session, bring extra clips to hold up your poster, and pack a few...seems captivated. Expertly navigate questions Answering questions is arguably the most difficult part ...very interested in his/her work, and would enjoy hearing the highlights so that you can appreciate the impact...

  32. Transferable Skills Guide: Managing a Team

    Type
    Blog Post
    ...transferable skills while working in the lab and by offering advice on areas where you might need some help...mine you didn’t think much about management skills during your scientific training. I never thought about...encourage questions. Unless you’re delegating a recurring task to a seasoned report, there should be questions...cases reports are not aware of issues until you bring them up. I wouldn’t go as far as saying that your...

  33. Viral Vectors 101: Voltage Indicators

    Type
    Blog Post
    ... subtypes, thanks to viral serotype specificity during delivery (more on this later). Chemigenetic sensors... at the single neuron level in vitro. In vivo monitoring has also been achieved with these sensors, but...their fluorescence, which ultimately led to the engineering of the second type of these GEVIs: a FP-fused...component, the chemical factor, of a naturally occurring voltage indicator. To give an example, a genetically...

  34. Of Myc and Men

    Type
    Blog Post
    ...not recognize the mouse or chicken homologs. By comparing the homologous sequences in these species, they...making the antibody more amenable to protein engineering. As a result, you can now find Myc1-9E10 in a...published a paper showing that the amino acids neighboring the Myc epitope influence 9E10’s affinity for...highly variable epitope recognition dependent on neighboring sequence context. Sci Signal 13. https://doi....

  35. Deep Dive: Fixing and Permeabilizing for Immunofluorescence

    Type
    Blog Post
    ...steps and outlined the considerations to be made during each step. Now that you know the basic principles...harsh - lipids and soluble proteins can be lost during fixation with these chemicals, which impacts sample...and attention.   * There are other methods for recovering epitope availability following cross-linking ...Immunofluorescence Staining. Methods in Molecular Biology. Springer New York, pp 299–311. https://doi.org/10.1007/...

  36. Plasmids 101: Choosing an Antibiotic Resistance Gene

    Type
    Blog Post
    ...plasmids, or express a gene contained in a plasmid. During the propagation and/or transformation process, ...never receive a copy of the plasmid or will lose it during division. These non-plasmid-containing bacteria...while amp only affects the cell wall synthesis step during cell division. This is nifty because the recovery...it will induce mutations in the host/plasmid DNA during selection if it’s not completely bound up. While...

  37. CRISPR 101: Making a Knock-In Cell Line

    Type
    Blog Post
    ...find out! Homology-directed repair and genome engineering The easiest way to make a knock-in cell line ...built-in repair pathway cells already have for repairing DNA double strand breaks – (HR). HR is the dominant..., H., Ren, S., Yu, S., Pan, H., et al. Methods favoring homology-directed repair choice in response to.... A. Enhanced homology-directed human genome engineering by controlled timing of CRISPR/Cas9 delivery....

  38. Minigenomes - a Safe Way to Study Dangerous Viruses Like the Ebola Virus

    Type
    Blog Post
    ...minigenome system Similar to other RNA viruses, EBOV brings its own RNA-dependent RNA polymerase into the infected...partially control expression levels of each gene by altering the amount transfected (see Technical Tips). Minigenome...changing the minigenome cis-acting sequences and monitoring effects on genome replication and transcription...strand RNA viruses: The power of reverse genetics. Springer Berlin Heidelberg, 2004. 1-41. PubMed PMID:15298166...

  39. Tips for Screening with Yeast Two Hybrid Systems

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    Blog Post
    ...eukaryotic transcriptional activation and cell biology. During this period, proteins were successfully expressed... own, but, if the two POI’s interact, this will bring the POI-TA fusion (commonly referred to as the “.... Lowered expression levels will increase the stringency of your binding. And most importantly: Independently...Ptashne, M. A eukaryotic transcriptional activator bearing the DNA specificity of a prokaryotic repressor....

  40. Fluorescent Proteins 101: Green Fluorescent Protein (GFP)

    Type
    Blog Post
    ...activity – it only requires the presence of oxygen during maturation. This meant that the protein could be...study of dynamic, physiological processes. Re-engineering GFP to increase its range of color and application...its sequence was elucidated, scientists began engineering new versions of GFP through mutagenesis in order...us how! And come back next week when we'll be featuring a guest blog post from Gal Haimovich of greenfluorescentblog...
Showing: 481 - 520 of 1006 results