...genes for drug discovery or to understand the intricacies of molecular pathways. These screens have gotten...diseases, as you can target your library to a significant degree using different types of AAV. Expression...into the genome, CRISPR may increase this rate. Specifically, portions of the AAV genome can integrate at...multi-plasmid system, where you have one AAV vector dedicated to expressing Cas9, while the other houses the...integration into CRISPR-induced DNA breaks. Nature Communications, 10(1). https://doi.org/10.1038/s41467-019-...

...efforts make a precise edit or insertion via a chemical modifier. We used these categories to see what... and the longevity of tools seen in Figure 3, indicates that the usefulness of popular CRISPR tools is...other countries, from Japan to Australia, are significant depositors and requestors. CRISPR tools are developed... into Addgene’s CRISPR data? Read our recent publication, “The Expanding Dissemination and Distribution...support for, this and previous Addgene CRISPR publications.  References and resources References Pyhtila...

...BLAST sequence databases from the dropdown menu. Typically, we use the default nucleotide database “Nucleotide...Organism box. This small piece of information can significantly reduce your wait time for blastn, blastp, and...” program for blastn so that you can extract practically every single reliable base pair from your results... the “Align two or more sequences” option to drastically reduce waiting time for search results. Optimizing...other labs speed up their plasmid and cloning verification steps and free up more time for using your plasmids...

...transforming a plasmid into the bacterium Agrobacterium tumeficans - a plant pathogenic species that stably integrates...Agrobacterium tDNA insertion likely precipitated domestication of sweet potatoes (Kyndt et al., 2015)! It sounds...selection markers (basta, hygromycin, kananamycin, tunicamycin) (Nakagawa et al., 2007). Alternatively, if you...plants: Lots of plants produce lots of seeds. I typically use 8-10 plants per construct I transform. More...National Academy of Sciences of the United States of America. National Academy of Sciences, 112(18), pp. 5844...

... of these techniques have in common? They all typically require secondary antibodies, frequently of the...in bacteria and extracted with common protein purification methods. Read on to learn more about nanobodies..., Alexa Fluor 568, or Alexa Fluor 647 before application to the sample. Staining with this multi-color...PubMed PMID: 29263082 Find plasmids from this publication at Addgene. 2. Bates, M., Böhning, M.I., Chug...antibody-containing hybridoma supernatant is only $40, significantly cheaper than commercially available antibodies...

...the Society for Advancement of Chicanos/Hispanics and Native Americans in Science’s (SACNAS) annual conference... written up in mSphere. A post-meeting survey indicated that the program was largely a success, but also... even more valuable program to Microbe 2020 in Chicago.  Finding our place in networking and mentoring... career scientists, students, and those from historically underrepresented groups. Through such mentoring...

...Why pseudotype? Lentiviruses and retroviruses typically target immune cells with their wild-type envelopes...out this review article published in Reviews in Medical Virology (Li et a., 2018).   Figure 2:...The glycoprotein will be incorporated into the replicating rabies virus envelope, travel retrogradely, and... Pseudotypes: Precious Tools to Improve Gene Modification of Hematopoietic Cells for Research and Gene...Restriction of Transsynaptic Tracing from Single, Genetically Targeted Neurons. Neuron 53:639–647 . https:/...

...addition, plasmids are typically prepared using endotoxin-free plasmid purification kits.     Figure...minimized by using an endotoxin-free plasmid purification protocol. To minimize the immunogenic properties... a concentration step prior to being frozen. Specifically, lentiviral particles are collected from the... Addgene's Data Hub Digital Droplet for AAV Quantification Viral Vectors 101: An Introduction to AAV References...

...Red arrows indicate a conversion directly catalyzed by the base editor. Black arrows indicate natural cellular...researchers can now select the best tool for their application from among many options.  Conclusion With these...engineered human uracil DNA glycosylase. Nature Communications, 15(1), 4897. https://doi.org/10.1038/s41467...via engineered uracil-DNA glycosylase. Nature Communications, 15(1), 6397. https://doi.org/10.1038/s41467...

... serotype, are particularly challenging. Their efficacy in any given system can be affected by any number... the Addgene repository can save researchers significant time and removes a variable from the lengthy ...available in the repository. GCaMP, a calcium indicator (biosensor, by Addgene’s categorization) was first...trends in our own repository data, giving us significant insight into which vectors are likely to be of...Waxmonsky, N. C., Swanson, L. E., & Fan, M. (2016). Practical Considerations for Using Pooled Lentiviral CRISPR...