...particle to infect a cell, it has to come into physical contact with that cell. Since there is a limited... by an anterograde AAV was shown to drop off drastically with lower titers. As you can see in figure 2...AAV-mediated gene expression in a target cell. Practical considerations If you have high quality AAV, you...Integration. Molecular therapy : the journal of the American Society of Gene Therapy. 2008;16(7):1189-1199.... Protocols for Work with Virus Watch Our AAV Purification Video ...

...imaging apoptosis in animals." Journal of the American Chemical Society (2019). PubMed PMID: 30821975. Additional...cell death" plays a pivotal role in an array of biological processes including development, the immune system...study apoptosis in vivo, scientists have used genetically encoded fluorescent executioner caspase reporters... reporter for imaging apoptosis in vivo." Cell chemical biology 23.7 (2016): 875-882. PubMed PMID: 27447051...

... Writing schedule tools How to write a lot: A practical guide by Paula J. Silvia  This book has tips for...app lets you track habits, like writing. It’s basically the equivalent of crossing off days on the calendar...suggestions Hemmngway makes (it wasn’t designed for technical writing), it’s great at flagging passive voice...jargon, this tool can help you avoid overusing technical terms when writing for a general audience. The...goal of the phrase, such as being cautious, being critical, or defining terms. The phrases in Phrasebank ...

...Collaboration and creativity are pivotal in writing publications/grants too. As described by several works (Wuchty...backgrounds.” This work outlines the need for clear communication, fair credit through authorship, and acceptance...remotely. The authors stress the fact that “miscommunication, a lack of leadership, and inappropriate tools...approaches can lead to frustration, delay of publication, or even the termination of a project.” Take ...setting. Check out the blog post “Fostering Communication in Remote Collaborations” to get more ideas....

... Connie Cepko (Harvard Medical School) , and Dr. George Church (Harvard Medical School) for taking the...environment for research is totally dependent on communication amongst the team so getting everyone talking...your department, and online. The Howard Hughes Medical Institute (HHMI) offers some great resources for...Scientists, including their "Making the Right Moves: A Practical Guide to Scientifıc Management for Postdocs and...

...North Carolina, Connie Cepko’s lab from Harvard Medical School, and Jonathan Ting’s lab from the Allen ...directions. I would like to thank them for their time, dedication, advice and encouragement. Why virus? Why did...educational materials, and provides viral vector technical service (details about virus production protocols...word out about this new resource and providing fantastically useful educational content on viral vectors....the impression that it took a lot of work and dedication to do this well ... because it did. So finally...

...remotely as an Addgene Science Communication Intern. Developing science communication skills, while also learning...panel about “Social Media for Science Communication” at the American Society for Cell Biology (ASCB) 2018...Twitter Learn about using Reddit for science communication ...

... the steps of gel purification and in vitro recombination. Across all modifications described in Trehan...effects of inactivating/activating mutations. In biomedical research, modeling patient mutations using SDM...mutations, large deletions/insertions require complicated, often costly in vitro assembly methods. A new...recombination machinery, which facilitates easy genetic modification by homologous recombination. In most recombineering...

...activate essentially any gene,” she says. The technical leap from mutating and deactivating a gene or ... and it consists of three components: 1. A nucleolytically inactive Cas9-VP64 fusion; 2. A sgRNA incorporating...distinct effector domains that work together synergistically, modeled after natural transcription activation...demonstrated in a paper in Cell that they could typically achieve 90-99% knockdown with minimal off-target...more about the rules.” He said that there is no theoretical limit on the number of genes that can be manipulated...

...improves upon them. Methods for studying autophagy Biochemical assays Long-lived protein degradation: This...radioactivity in the culture supernatant (this is indicative of protein degradation). To control for protein...’s slow and only measures bulk autophagy. Morphological assays Electron microscopy: Electron microscopy...studying autophagy. This method relies on the identification of autophagic structures based on morphology...available from Addgene. References 1. Rosado, C., Mijaljica, D., Hatzinisiriou, I., Prescott, M., & Devenish...

...time-consuming aspects of traditional cloning is the identification of clones that actually contain your insert...enzyme-mediated disruption of ccdB, leading to positive identification of desired clones.   ...Becomes a powerful...ccdB and therefore would die. This procedure dramatically reduces the number of clones that do not contain...which can then be used for downstream cloning applications. While these two strains ultimately perform ...and ccdB Survival strains have been developed specifically with ccdB-containing vectors in mind, any plasmid...

... year, narrow by article type (review, patent, clinical study, research article etc). There are also numerous...looking for a particular protocol or are trying to replicate an experiment, the methods section probably won...conclusions. In any case, the reader should give a critical eye to all results in the relation to how they...is science’s way of validating work prior to publication, it doesn’t mean that there are no mistakes or... biochemistry instructor in the Department of Biological Sciences at Louisiana State University. She is...

...including RNPs for indel formation via NHEJ, precise modification with Homology Directed Repair (HDR), CRISPR ...potential to be used for other types of CRISPR applications. Let’s take a look at the key components of ... which means they are non-infectious and non-replicative. Nanoblades are produced by transfecting the ...cytoplasm or pronucleus of the zygote which is both technically challenging and can negatively impact zygote ...with Cas9-sgRNA ribonucleoproteins." Nature communications 10.1 (2019): 45. PubMed PMID: 30604748. PubMed...

...when it comes to targeting precision and practical application. If the CRISPR/Cas9 system is not effective...but directly incorporating very long ncRNAs is impractical. Instead, tags or stop elements can be inserted...can cleave single-stranded RNA using gRNAs to specifically edit or degrade targeted lncRNAs. The Cas14 ...lncRNAs with high specificity, particularly in applications requiring smaller delivery systems (Zibitt et...

...biologists and computer scientists learning and communicating with one another.      Currently our main focus...our worms.  We are on the lookout for genome modifications that do not exist in the CGC library but that...makeup of the bacterial species in the gut of the domesticated housecat, and seeing how that makeup differs...cats with the goal of optimizing the health of domesticated cats and learning about feline microbiomes. ...and the reagents are too expensive to make it practical - but we have members looking at how to re-use...

...Chavez, researchers at the Wyss Institute for Biologically Inspired Engineering. CRISPR/Cas9 technology... addition to its utility in making targeted modifications to DNA, Cas9 can be reprogrammed to serve as... a gene of interest, they have been shown to dramatically alter levels of expression. Cas9’s ability to...Orthogonal gene knockout and activation with a catalytically active Cas9 nuclease." Nature biotechnology ...Options for CRISPR Genotyping Cas9 Activators: A Practical Guide Additional Resources on Addgene.org Check...

...The vast majority of bacteria are undomesticated which limits the tools scientists can use to study them...the genome and is propagated during normal DNA replication and cell division. Mobile-CRISPRi does not disrupt...single-gene knockdown experiments. Mobile-CRISPRi applications Mobile-CRISPRi expands CRISPRi to non-model ...Elizabeth A., and Pamela A. Silver. "Harnessing undomesticated life." Nature microbiology 4.2 (2019): 212....PMC6424567. Peters, Joseph E. "Tn7." Mobile DNA III. American Society of Microbiology, 2015. 647-667. PubMed...

...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...

...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-...

...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...