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  1. How Addgene Creates Content

    Type
    Blog Post
    ... ideas come from social media conversations, conferences, and questions our scientific support staff receives...line with Addgene style.        Fig. 2: Reference image (left) provided to our graphic designer ...constantly reviewing the resources we currently offer to identify gaps and add to them as techniques evolve...
  2. Giving gRNAs a Facelift - Synthetic and Beyond

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    Blog Post
    ...experiment moving. Happy editing! References and Resources References Hendel, A., Bak, R., Clark, J. et...used in CRISPR-Cas9 experiments – the “gRNA” – suffers from the same issues as your experimental RNA. ...
  3. A Deep Dive into BioBuilder

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    Blog Post
    ...’s classrooms,” BioBuilder answers the call by offering a variety of educational programs all focused ...opened in August 2017.  This flexible teaching space offers STEM programming, workshops, and customized programs...point into further STEM exploration.  Further offerings for both STEM teachers and STEM-loving students...
  4. Multiplex Genome Editing with CRISPR-Cpf1

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    Blog Post
    ...can lead to variable expression levels due to differences in copy number.  And there are the usual transfection...Method Delivery Method Advantages Disadvantages References Yamamoto Lab Golden Gate Assembly transfection...multiplexing, leave them in the comments below.        References 1. Zetsche et al. "Multiplex Gene Editing by ...
  5. Viral Vectors 101: The AAV Retrograde Serotype

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    Blog Post
    ...that pathways projecting to the inferior colliculus (IC) are differentially transduced when using the retrograde...significantly updated in 2023 by Jason Nasse.  References 1. Tervo, D. Gowanlock R., et al. "A designer...
  6. REPLACR Mutagenesis: Replacing In Vitro Recombination Methods

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    Blog Post
    ...synthesis, and multifragment assembly methods are preferred. No such restriction applies to deletions; using...common methods for site directed mutagenesis   References 1. Trehan, Ashutosh, et al. (2016). “REPLACR-... on Addgene.org Browse Our Molecular Biology Reference Pages Choose Your Perfect Plasmid Backbone Find...
  7. Viral Vectors 101: Viruses as Biological Tools

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    Blog Post
    ...nature, viruses infect cells and organisms (which we refer to as hosts) via viral particles, the functional...deliver cargo, it’s important to understand their differences to choose the best one for your experiment. ...Here to Find Viral Vector Resources at Addgene! References and Resources 1. Roe T, et al. “Integration of...
  8. Developing a cold tolerant E. coli using specialized chaperones

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    Blog Post
    ... of interest). Mesophilic microbes are highly preferable (and successful) as host systems since their ...to guess. Think about it like winter tires in different conditions: the more flexible rubber of winter...reagent repositories and the myriad facilities offered by them which help foster a more productive and...
  9. Back to Bacteria: CRISPR gRNA Multiplexing Using tRNAs

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    Blog Post
    ...expression, PTGs will likely be adaptable to many different biological systems. Although Xie et. al characterized...in plants, PTGs will likely work well in many different biological systems. Size is also a factor; the...technology, leave them in the comments below.  References Boosting CRISPR/Cas9 multiplex editing capability...
  10. RaPID Detection of RNA-protein Interactions

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    Blog Post
    ...affect protein binding patterns. Hereditary Hyperferritinemia-Cataract Syndrome (HHCS), a disease characterized...the Iron Response Element (IRE) RNA motif of the ferritin light chain RNA transcript. IRE motifs are bound...The RaPID plasmids are available from Addgene! References Ramanathan, M., Majzoub, K., Rao, D.S., Neela...
  11. RANbodies: Reporter Nanobody Fusions

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    Blog Post
    ...difficult for an individual lab to produce - Hybridomas offer an alternative and renewable source of monoclonal... their use. Each RANbody is detectable by two different tags: a reporter and an HA tag. The reporter comes... link to its sequence in the comments below. References Sanes, J.R., & Yamagata, M. (2018). Reporter-...
  12. Visualizing Protein Turnover In Situ

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    Blog Post
    ...overexpressed from a plasmid but overexpression may interfere with the system. Another method for studying protein...whether or not the tags influence behavior. A conference and a breakthrough! During my graduate studies...you can contact her at [email protected]. References 1. Dieterich, Daniela C., et al. "In situ visualization...
  13. Supporting Rare Disease Research with the 2020 BeHEARD Awards

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    Blog Post
    ...newly independent faculty member studying sex differences in thoracic aortic disease,” Prakash says. In..., the lab hopes to capture the signalosome of different receptors upon stimulation. “To study cellular... Jennifer Tsang...
  14. Antibodies 101: Affinity Reagents

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    Blog Post
    ...bispecific, meaning that each scFv can target a different antigen; binding of each can be either simultaneous... a scientist looking for that perfect fit!  References and Resources Additional resources on the Addgene...AddgeneBrowse our recombinant antibody plasmid collection References Grumezescu, A. M. (2018). Drug targeting and ...
  15. Choosing the B(right)est Fluorescent Protein: Aggregation Tendency

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    Blog Post
    ...dTomato) that promote interactions between the FPs interfere with native interactions, preventing proper localization...papers should be used with caution, since the difference between studies is substantial (e.g, the poor...conclude, we recommend that you try a couple of different fluorescent proteins when protein tagging is the...
  16. Enhancing CRISPR Targeting Specificity with eSpCas9, SpCas9-HF1, & HypaCas9

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    Blog Post
    ...mutations). Although the precise mutations were different than those found in eSpCas9, the effect on specificity...and/or therapeutic settings. Will combining the different mutations found in each of these great tools further...continued improvements in genome engineering! References 1. Slaymaker, Ian M., et al. "Rationally engineered...
  17. New Tool for Lineage Tracing: The ClonTracer Library

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    Blog Post
    ...replicates would have different labels - i.e. they would stochastically arise from different cells throughout...development of combination therapies.         References 1. Hyo-eun, C. Bhang, et al. "Studying clonal...
  18. Plasmids 101: Cre-lox

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    Blog Post
    ...because it “causes recombination” (although later referred to as the “cyclization recombinase”), is a 38 ...same type, but are not cross-compatible. Using different lox site variants allows Cre to catalyze more ...cre-dependent techniques?  Let us know in the comments! References 1. Bacteriophage P1 site-specific recombination...
  19. Adeno Associated Virus (AAV) for Cell and Gene Therapy

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    Blog Post
    ...their mutations corrected, and the T-cells were transferred back into the infants. Almost all patients have...population. These pre-existing antibodies can interfere with virus-cell interactions and seriously hinder... cure research and science communication.     References 1. Gaspar, H. B., S. Howe, and A. J. Thrasher...
  20. Plasmids for Endogenous Gene Tagging in Human Cells

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    Blog Post
    ...methods flood the cell with protein, which can both interfere with a cell’s normal function and result in a ...portal. The plasmids should be functional in many different human cell types, and while most of the genes ...Allen Institute website at alleninstitute.org. References 1. Roberts, Brock, et al. "Systematic gene tagging...
  21. CRISPR in the Clinic

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    Blog Post
    ...updated in 2023 by Susanna Stroik.  References and Resources References Yuxuan Wu et al. "Correction of a...frequency isn’t high enough to make a physiological difference, then the treatment will not work in a meaningful...
  22. High Complexity Golden Gate Assembly with NEB’s DAD

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    Blog Post
    ...base or 4-base overhangs, and with a variety of different enzymes and cycling conditions (see Table 1). ... cycling 3-base  PaqCl®, 1x T4 DNA Ligase Buffer 37-16 °C cycling 4-base PaqCl, 1x NEBridge... and don’t forget to ask DAD for some help! References Potapov, V., Ong, J. L., Kucera, R. B., Langhorst...
  23. Viral Vectors 101: Virus Safety

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    Blog Post
    ...dividing up the essential components of a virus into different plasmids. For a retroviral system this looks like... like the genetic cargo in one plasmid (the transfer plasmid), the envelope (Env) gene on a separate plasmid...Env, Gag, and Pol genes are not encoded in the transfer plasmid, the component that physically integrates...
  24. Hot Plasmids: Winter 2025

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    Blog Post
    ... tomato plants that inherited an edited gene to confer resistance to the herbicide chlorsulfuron (Homo...want a negative-going GEVI that can be used in different illumination regimes with fast activation kinetics... a YFP-tagged G protein and a NanoLuc® (Nluc) Luciferase-fused detector that specifically binds to active...
  25. Magnetic Control of Proteins: More than a Dream

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    Blog Post
    ...Find the MagLOV plasmid here!   References and resources References Duret, G., Polali, S., Anderson,...just be an artifact and eliminate sources of interference, but the result persisted. (Even better, multiple...
  26. Light Sheet Fluorescence Microscopy

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    Blog Post
    ...spinal tract, CST, spinal cord and cranial nerve afferents labelled with a AAV8 expressing eGFP under the...sample and captured the reflected light using a different objective lens oriented perpendicular to the plane...Find plasmids from the Tsoulfas lab here.     References: H. Siedentopf, and R,. Zsigmondy.  (1902) Ann...
  27. Transgenic Organisms, Cas9 Gene Drives, and Appropriate Safeguards

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    Blog Post
    ... lines with compound autosomes are completely infertile when mated to wild fruit flies6. It's also worth...information about gene drives can be found in the referenced publications. Thank you to our guest blogger...information, visit www.sculptingevolution.org. References: Burt, A. Site-specific selfish genes as tools...
  28. Interview: Nicola Patron on Plant Synthetic Biology, MoClo, and More

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    Blog Post
    ...explains it, her work has always been focused on gene transfer, from transgenes to plants, chloroplast to the...anymore. The kit contains sequences for several different fluorescent proteins. Visualizing fluorescence...for those who go to work in cancer biology.  References: A Golden Gate Modular Cloning Toolbox for Plants...
  29. Drew Endy Introduces the Biobrick Public Agreement Plasmid Collection

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    Blog Post
    ...the physical DNA samples, for when that's the preferred mode of receiving parts." "We have found an amazing...functions, which we know and love. Boolean is in reference to George Boole who in 1854 was thinking about...tell us readers, what do you want to make?   References: Jerome Bonnet et al. "Amplifying genetic logic...
  30. Tips for arabidopsis transformation

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    Blog Post
    ...plant pathogenic species that stably integrates transfer DNA (tDNA) into the genomes of the plants it attacks... (Nakagawa et al., 2007). Alternatively, if you prefer Golden Gate assembly, there are a series of plasmids...countless transgenic plants in the process.     References 1. Emami, S., Yee, M. and Dinneny, J. R. (2013...
  31. Capturing a Moment with Marc Zimmer’s “The State of Science”

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    Blog Post
    ...journal editorials, science social media, and conference workshops for years. That is, presumably, why...these fields, and seeing names like Feng Zhang, Jennifer Doudna, Karl Deisseroth, and Ed Boyden crop up...scientist who enjoys learning a little bit about many different things (part of the reason Addgene has been such...
  32. Isolating Genomic Regions of Interest with the CRISPR System

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    Blog Post
    ... adaptations of this system have been made in different fields of study. In addition to the identification...purification with an anti-Cas9 antibody. These different tag systems and affinity purification schemes ...please also see our interview with Dr. Fujii. References Campbell, Amy E., et al. "NuRD and CAF-1-mediated...
  33. Better Dyeing Through Chemistry & Small Molecule Fluorophores

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    Blog Post
    ... lipoic acid ligase, biotin ligase, phosphopantetheinyltransferase) – These enzymes catalyze the covalent... We continue to develop derivatives that have different spectral properties, exhibit high fluorogenicity...modalities. Follow him on twitter @rhodamine110.   References 1. Liu, Zhe, Luke D. Lavis, and Eric Betzig. ...
  34. A Career in Grants: How To Become a Grant Professional

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    Blog Post
    ...development, and more; our work touched on many different issues, and our program didn’t stay in one place...management, and program evaluation are highly transferable. Use your experience and your passion(s) to ...coach you. There are many grant consultants who offer grant training, mentoring and coaching services,...
  35. Neuronal labeling with Spaghetti Monster

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    Blog Post
    ...tracking experiments (Rizzo, 2009). With many different FPs that emit across the visible spectrum, researchers...antibodies. The “spaghetti monster” name is a reference to the deity of the social movement, the church... to the expanded use of smFPs in the future. References 1. Viswanathan, Sarada, et al. "High-performance...
  36. Rett Syndrome: A History of Research and Therapeutic Outlooks

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    Blog Post
    ...these discoveries, scientists have studied how different parts of MECP2 influence Rett Syndrome. Zoghbi...varying degrees determine the clinical severity of different MECP2 mutations. More recently, the Bird lab found..., Rett databases, and other helpful tools.  References: Amir RE, Van den Veyver IB, Wan M, Tran CQ, ...
  37. The PAM Requirement and Expanding CRISPR Beyond SpCas9

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    Blog Post
    .... Note: Jennifer Tsang and Mary Gearing contributed to the update of this post. References Abudayyeh...2016). Type VI CRISPR systems, which target RNA, offer additional targeting flexibility. Cas13a (formerly...
  38. CRISPR-mediated Plant Base Editors

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    Blog Post
    ...0-5%. This blog post covers a brief review of different basic BE technologies and their adaptation for...Nickase Cas9 (D10A) Deaminase PAM requirements Reference CBE SpCas9 rAPOBEC1 NGG 6 SpCas9 Target-AID...improvement in the laboratory of Yinong Yang at PSU.   References 1. Komor, Alexis C., et al. "Programmable editing...
  39. Tips for Writing a Good Cover Letter

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    Blog Post
    ...can use your cover letter to explain why your transferable skills make you a good candidate for the position...experience or skills, often by pointing to your transferable skills. In the above example, if you don’t have...experience, having given more than 20 talks at conferences and taught for five years at [name of the university...
  40. Plasmids 101: E. coli Strains for Protein Expression

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    Blog Post
    ...below, certain specialized strains are available to confer greater transcriptional control, assist with proper...pLys comes in two flavors—pLysS and pLysE—the difference being that the latter provides tighter control...minimal media supplemented with M9 salts may be preferable if the protein product is secreted to the medium...
  41. Plasmids 101: The Promoter Region – Let's Go!

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    Blog Post
    ...host organism. Because transcription machinery differs between cell types or organisms, promoters must...eukaryotes and prokaryotes We have included two reference tables below listing some of the most common bacterial...Mammalian Vectors Read about Reporter Gene like Luciferase and GFP Resources on Addgene.org Find Plasmids...
  42. SARS-CoV-2/COVID-19 Detection Methods Based on CRISPR/Cas

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    Blog Post
    ...Cas12a, Cas13a does not require strict sequence preferences at the target site, while Cas12a requires a PAM...femto-aptomolar range. For more details on the differences between these methods, check out this table.  ...can follow her on Twitter @shra_krishna.       References Azhar, M., Phutela, R., Ansari, A. H., Sinha,...
  43. Harnessing Bacterial Toxins for Allelic Exchange

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    Blog Post
    ...leaves a “scar” in the chromosome, sometimes interfering with expression of surrounding genes. However...Gram-negative bacteria become resistant to antibiotics! References Espah Borujeni, Amin, Anirudh S. Channarasappa...for allelic exchange Read our molecular biology reference Watch molecular biology protocol videos ...
  44. CRISPR 101: Validating Your Genome Edit

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    Blog Post
    ...dice on a new gRNA in the future! References and Resources References Eva K. Brinkman, Tao Chen, Mario ... frame editing frequency is 50%, then you could infer that approximately 25% of cells are likely -/-, ...
  45. Quick Guide to Near-Infrared Fluorescent Proteins

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    Blog Post
    ...recent research @daria_gift_shch.     References and resources References Shemetov, A.A., Oliinyk, O.S. & ...the monomeric state of a probe leads to less interference with the oligomeric state of a fusion partner...
  46. Targeting HIV-1 with CRISPR: Shock and Kill or Cut it Out?

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    Blog Post
    ...binding sites increased activation of HIV promoter-luciferase constructs. In multiple HIV-1 latent T cell lines...the HIV-1 copy number, although the efficiency differed from 48-100% between strains. In a similar experiment...for some of the world’s most common diseases. References Zhang Y, Yin C, Zhang T, Li F, Yang W, Kaminski...
  47. How to Negotiate a Successful Lab Start Up Budget

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    Blog Post
    ...on unnecessary materials, or even a loss of the offer all together. The problem is that the discussions...specialized equipment, try finding institutes that offer shared equipment that you can use at a lower cost...turned down a faculty position because none of the offering institutes were able to provide the resources ...
  48. Hot Plasmids: Spring 2025

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    Blog Post
    ...control of one of seven enhancers specific for different interneuron populations. The enhancers span all...(LAMP5) — as well as cholinergic (choline acetyltransferase, ChAT) neurons. The authors characterized ...effort to develop molecular tools for studying different brain cell types. Look for more of these interneuron-targeting...
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