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TypeBlog PostPublishedOct. 17, 2017, 12:55 p.m....crystallographer working with Jane Richardson, provided an initial solution to this problem with the development...the surface of the protein where the nascent polypeptide emerges from the exit channel. He then said...allow the complementary base pairing between the guide RNA and the target DNA strand, and then activate...User Resources on Addgene.org Visit Our CRISPR Guide Page CRISPR/Cas9 History Browse All CRISPR Plasmids...
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Hot Plasmids: Summer 2025
TypeBlog PostPublishedAug. 28, 2025, 1:15 p.m....fluorescent proteins just haven’t met your needs, consider trying mGold2s or mGold2t for your next experiments...TnpB is directed to genomic targets using an RNA guide (known as an omega RNA, or ωRNA) (Figure 2), but...Weiss, T., et al. (2025). Viral delivery of an RNA-guided genome editor for transgene-free germline editing... Mike Lacy Whether you’re planning on lounging poolside or swimming laps, Addgene has pooled libraries... -
Degrading DNA with Cascade-Cas3
TypeBlog PostPublishedFeb. 11, 2025, 2:15 p.m....powerful than it is. The nuclease domain of Cas3 is considered weak, especially compared to the power of Cas9...Repetitive DNA reeling by the Cascade-Cas3 complex in nucleotide unwinding steps. Molecular Cell, 70(3), 385-394...Liao, M., & Ke, A. (2018). Structure basis for RNA-guided DNA degradation by Cascade and Cas3. Science, 361...CRISPR-Transposons Additional resources on Addgene.org CRISPR guide Cascade-Cas3 Plasmid collection ... -
Viral Vectors 101: Producing Your rAAV
TypeBlog PostPublishedJuly 16, 2024, 1:15 p.m....There are several facilities nationwide where you can obtain excellent quality, high titer rAAV (including...certain in vitro and in vivo applications, you can consider using unpurified AAV from the supernatant (Goodwin... in precision and variability, qPCR is the more widely used method due to the accessibility of instruments...101 series! This post was originally written by Didem Goz Ayturk and updated by Ina Ersing in August 2023... -
Tips for a 1st time AAV user (by a Rookie AAV user)
TypeBlog PostPublishedOct. 23, 2018, 11:49 a.m....plasmids to produce AAV: 1) the RepCap plasmid which provides the AAV replication (rep) and capsid (cap) genes...primarily found in the culture media, you should consider performing PEG precipitation. Instead of sonicating... details. Pro tips Watch this AAV purification video! It didn’t exist when I did my first iodixanol gradient... gradient purification, but I wish it had. The video gives a brief overview of how iodixanol purifications... -
10 Basic tips for mammalian cell culture
TypeBlog PostPublishedJuly 12, 2018, 1:09 p.m....of contamination for your experiments, the bright side is that you also have the power to take the added...working with so that cross-contamination can be identified. Even if some mammalian cells lines have similar...check under the light microscope will help you identify bacterial and yeast contaminants. Set up a routine...will help you avoid cross-contamination through accidental mixing and will also help you keep good records... -
Sharing Your Lab Protocols: Using Apps to Save Time & Track Your Experiments
TypeBlog PostPublishedSept. 23, 2014, 3:05 p.m....After learning of it's continued popularity, I decided this protocol would be a worthy first contribution...protocols. After my introduction to protocols.io, I decided to research what other online (and mobile) tools...www.addgene.org/ If you’re working with plasmids and you decide to share them using Addgene’s repository, you can...and allows scientists to contribute protocols or provide links to protocols on their own websites. We're... -
Interview: Nicola Patron on Plant Synthetic Biology, MoClo, and More
TypeBlog PostPublishedJune 12, 2014, 3:30 p.m....wanted to expand the toolkit. We have included a wider range of terminators than are commonly used as we...rusts and we are collecting sequences that will provide a similar resource for those involved in monocot... really and we’ll be producing an instructional video soon to make it easier for people to use the Golden...think plant biology is massively underfunded considering how important it is. I would really like to reach... -
Drew Endy Introduces the Biobrick Public Agreement Plasmid Collection
TypeBlog PostPublishedDec. 12, 2013, 3:35 p.m....DNA systems capable of storing data and computing inside living cells. He's also a co-founder of the BioBricks...Champion of Change, a program created as part of President Barack Obama's Winning the Future Initiative. ...proteins. That's central to everything that goes on inside a cell. Until we get a handle on precise, forward...promise that we could instead have things like peptide or RNA aptamers that act as genetically encoded... -
CRISPR Antimicrobials
TypeBlog PostPublishedMay 3, 2016, 2:30 p.m....less potential to become resistant to? CRISPR may provide a method for doing just that. While challenges ...problem. Above all, we need new antimicrobials, ideally ones for which the development of resistance will... antimicrobials using efficiently-delivered RNA-guided nucleases.” Nature Biotechnol. 32(11): 1141-5. ...CRISPR Plasmids for Your Research Read Our CRISPR Guide Pages Find CRISPR gRNA Design Tools ... -
Getting to Know Expansion Microscopy
TypeBlog PostPublishedJune 20, 2023, 1:15 p.m....Microscopy. What is Expansion Microscopy? ExM provides a straightforward answer to the space problem....minimized through tweaks to these steps. But even under ideal polymerization and digestion conditions, loss of...Bournonville L, et al (2021) Expansion microscopy provides new insights into the cytoskeleton of malaria ...Sommer C, Rizzoli SO, Danzl JG (2019) A practical guide to optimization in X10 expansion microscopy. Nat... -
Capturing a Moment with Marc Zimmer’s “The State of Science”
TypeBlog PostPublishedJuly 30, 2020, 1:15 p.m....inclusion of some interesting case studies, and Zimmer provides several. In Part Three of the book, called simply... and her essential-but-unlauded role in the identification of DNA’s double helix. Perhaps to the dismay...dismay of middle-aged scientists, the identification of Green Fluorescent Protein (GFP) and its development...(for funding, etc.) with that produced by the confident and flamboyant Watsons.” In Part Four, “New Science... -
Tips for Screening with Yeast Two Hybrid Systems
TypeBlog PostPublishedOct. 22, 2015, 2:30 p.m....numbers, but the latter eliminates the step of identifying prey proteins after screening. It is also possible...activation. Including a prey-only control also provides a baseline level of reporter activation. Vary ... And most importantly: Independently validate identified binding partners with other techniques! Minimizing... While this may be difficult to resolve for a wide range of prey proteins, there are now a number of... -
Viral Vectors 101: Pseudotyping
TypeBlog PostPublishedApril 20, 2021, 2:30 p.m....pseudotyping allows these viruses to infect neurons. Besides altering the host tropism of your virus, there ... as a retrograde tracer so that scientists can identify the neurons that project to a given brain region... resources on the Addgene blog: Read the Quick Guide to All Things Lentivirus Get tips for titering your...Resources on Addgene.org: Check out the lentivirus guide Find ready-to-use lentiviral preps at Addgene ... -
Plasmids 101: Plasmid Incompatibility
TypeBlog PostPublishedMarch 31, 2020, 1:15 p.m....theory for plasmid incompatibility is due to an identification issue: the CBPs in the cell can’t tell the ... lost from the cell at a higher rate than the coresident plasmid. Asymmetric plasmid loss occurs for several...based on sequence data. However, if these do not provide clarity, a wet lab transformation experiment will...Genomics of microbial plasmids: classification and identification based on replication and transfer systems and... -
Rett Syndrome: A History of Research and Therapeutic Outlooks
TypeBlog PostPublishedNov. 19, 2020, 2:15 p.m....that MECP2 binds to methylated CpG dinucleotides and CAC trinucleotides during neuronal maturation (Tillotson...Sinnamon et al., 2020). They introduced a mouse MECP2 guide RNA and a hyperactive version of the human RNA base...fine-tuning dosage so that enough wild-type MECP2 is made inside the cells, but this is exciting progress in tackling... -
CRISPR-mediated Plant Base Editors
TypeBlog PostPublishedJan. 3, 2019, 1:35 p.m.... defective gene, Xm, the sequence of which is identical to the correct gene, Xw, except for a single base...vectors have two Bsa1 sites to clone the single guide RNA (protospacer+scaffold) in a pol-III (OsU3) promoter... in a ‘GC’ sequence context. If this cannot be avoided, select the target AID version. Expect very low...PMC4873371. 2. Nishida, Keiji, et al. "Targeted nucleotide editing using hybrid prokaryotic and vertebrate... -
Hot Plasmids Spring 2024
TypeBlog PostPublishedMay 23, 2024, 1:15 p.m.... Adamson Lab recently used a CRISPRi screen to identify additional host factors impacting prime editing...subthreshold voltage fluctuations, while having high fidelity during action potentials of fast-spiking neurons... combination with miR124T targeting sequences, provides a simple yet valuable tool for microglia research...2023). Simple and Highly Specific Targeting of Resident Microglia with Adeno-Associated Virus. bioRxiv... -
Plasmids 101: E. coli Strains for Protein Expression
TypeBlog PostPublishedFeb. 10, 2015, 3:06 p.m....growth phase are alive and healthy, which makes them ideal for protein expression. If you wait too long to ...and pLysE—the difference being that the latter provides tighter control of basal expression. What if I... growth media, or if your plasmid is low-copy, consider reducing the antibiotic concentration. Growth ...which contain hydrophobic regions or multiple disulfide bonds may aggregate and become insoluble. These... -
Four Ways to Package Transgenes That Exceed the Size Limit of Adeno-associated Virus
TypeBlog PostPublishedSept. 1, 2020, 1:15 p.m....First, some history. In 2008, the Auricchio lab decided to solve the issue of “tiny vector, big transgene...of expressing a truncated protein. This can be avoided by excluding a stop codon from donor A in any of... to be reconstituted, there is currently no one ideal split vector system. Overall the efficiency of transgene...Resources on Addgene.org: Check out the Addgene AAV guide Find ready-to-use AAV preps from Addgene's viral...