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...In Addgene’s early days, we would ‘spot-check’ critical regions of plasmids deposited with us via Sanger...development teams, frequent updates allowed us to scale our lab work as we grew. As the number of deposited...us to switch between executing our pipeline on a local computer or in a cloud environment. Given the throughput...jumping back and forth between running the pipeline locally and in the cloud. Creating a bioinformatics workflow...script file (containing our new process) on our local computer. Eventually, our workflow would handle ...

...experimental design and its potential for clinical applications. Specifically, efficacy rates of Cas7-11 were reported...current knowledge. The Abudayyeh-Gootenberg lab (also called the AbuGoot lab), a joint lab run by Omar Abudayyeh...originally suspected that this new Cas7-11 might be phylogenetically classified as the more complex Class I systems...switched up the serine integrase for one which can enzymatically take a genetic sequence and integrate it into...

...JAT, Callaway EM (2007) Monosynaptic Restriction of Transsynaptic Tracing from Single, Genetically Targeted...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:...6178.1997 Wickersham IR, Finke S, Conzelmann K-K, Callaway EM (2006) Retrograde neuronal tracing with a deletion-mutant...

...studying biochemistry at the University of Southern California. He is also an intern at the Yang Lab of the ...transposon-encoded CRISPR/Cas systems in bacteria, specifically on optimizing strains with low efficiency.   ...of Sciences. He joined Shanghai Institutes for Biological Sciences in 2000 and became a professor in Synthetic...reprogramming of cells to understand the genetic and biochemical mechanisms behind their distinct phenotypes and...generation of engineered cells for green chemistry and biomedical applications.   Yiwen Zhang entered the Sheng...

...administered systemically (retro-orbitally, RO; or intracerebroventricularly, ICV) or locally (stereotaxically...developed, funded, and shared has no doubt changed drastically over the past few decades. But no matter how ...and transgenic mouse lines for genetic access to cortical cell types. Cell. https://doi.org/10.1016/j.cell...

... an exciting class of compact genome editors, typically only around 400 amino acids in size. To give a...target DNA, TnpBs also need a short sequence motif called a TAM (transposon-associated motif), which is positioned...familiar targeting mechanism. Small RNA elements called right-end RNA (reRNA) or omega RNA (ωRNA) guide... a single experiment (Karmakar et al., 2024). Practical Guide: Using pK-TnpB1 and pK-TnpB2 Vectors The...The process for cloning the 20-nt guide RNA is identical for both vectors. Here, we provide a step-by-step...

...allows for directional assembly of DNA sequences, called modules or parts. Assembly is dictated by four-...for fresh bacteria. Performing your Assemblies Calculate your volumes As you progress through the MoClo...increase your ligation efficiency, we recommend you calculate the right volume of each plasmid according to ...the equation in Figure 3.    Figure 3: Calculation for the plasmid volume for MoClo experiments....high-throughput, we recommend transforming into chemically competent cells in a 96-well PCR plate. Always...

...production of viral vectors typically involves shuttle vectors, also sometimes called transfer vectors. The ...antibiotic works. Another example is Zeocin, a DNA intercalator, which affects all kingdoms of life. Of course...host range plasmids shuttle vectors? While it’s technically accurate to say that shuttle vectors have a “... “broad host range,” these terms are typically used in different (though overlapping) ways. Broad host...

...FRET during a journal club, my guitarist brain automatically thought about the raised element found on the...interactions in living cells using a microscope. Historically, molecular interactions were detected by indirect...know or interact with each other. FRET reduced the scale of our perception about molecular interactions. ...Periasamy defined FRET as "a distance-dependent physical process by which energy is transferred non-radiatively...FRET) microscopy imaging of live cell protein localizations. J Cell Biol. 2003 Mar 3;160(5):629-33. ...

... RT-qPCR tests to diagnose COVID-19 and some serological tests to determine past exposure. However, the...rapid detection.  Highlights: Cas13a is not catalytically active in the presence of two or more mismatches... copies RNA targets in a 40 min incubation. A scalable, easy-to-deploy, protocol for Cas13-based detection...available and less expensive. Instead of using the typical PCR machines, the authors use affordable, field-ready..., Acosta-Alvear, D., & Wilson, M. Z. (2020). A Scalable, Easy-to-Deploy, Protocol for Cas13-Based Detection...

...set of transferable skills, coupled with your technical skills, can open new career opportunities for ...conflict can also be good. In science, a reviewer’s critical comments on your manuscript is a form of (hopefully...scientist. And some amount of conflict is actually critical in the business world. Companies and organizations...aspects of customer support each team “owns” is critical for day-to-day operations and keeping our customers...especially people with whom you strongly disagree, is a critical component of conflict resolution. Having empathy...

...insights into the complicated physical organization of the brain and other biological systems. Interested in ...deletions are defined using 2 loxP variants (only identical loxP sites can mediate recombination.) The deletions... preventing further recombination. Brainbow-1.1 scales up the system to four fluorescent proteins, with...homeostasis results from neutral competition between symmetrically dividing Lgr5 stem cells. Snippert HJ, van der...

...without their problems, but much genetic and physiological data have been accumulated over the years using...check out our Mouse Modeling blog posts: Part 1: Genetically Engineered Mice and Part 2: Breeding and Crossing...into the oviduct of pregnant females, a technique called Genome editing via Oviductal Nucleic Acids Delivery...small mutations with HDR, when it comes to larger scale genome editing, such as replacement of a mouse gene...nice to your microinjectionist, who has the strategically important job of delivering the payload! Last...

...lab, or tissue culture as it is commonly called, is a critical tool for many scientists. Mammalian cells...AAV preparation. For detailed instructions and calculations on how to prepare a plasmid stock used to generate...cells provide scientists with the means to study biological processes on the cellular level instead of having...

...completing her PhD in Chemistry and Chemical Engineering with Harry Gray at Caltech, Crystal moved on to a Postdoc...Investigator III at Novartis, a multinational pharmaceutical company based in Switzerland. After completing...scientific teams at the Novartis Institutes for BioMedical Research to develop novel biologics for the treatment...

...Addgene podcast, we sit down with Alex Chavez, a Clinical Fellow in Pathology at Masachusetts General Hospital...Church and Collins labs at the Wyss institute for Biologically Inspired Engineering. Alex is the first MD PhD...and explains why he loved the first 2 years of medical school. 7:05 - 10:55 : Alex's focus during graduate...postdoc 12:01 - 13:21 : The Wyss Institute for Biologically Inspired Engineering and why Alex decided to...

...use as a genetic engineering tool is frequently called recombineering - short for homologous recombination-mediated...insert site (Figure 2, top).  These primers are typically ~70 nucleotides long (20 nucleotides that anneal...fragments, such as gene replacements and tags.  The typical frequency of recombinants is 1 positive clone out...efficiency than the other and you’ll find out empirically which is the lagging strand. ssDNA substrate ...below).  Controlling expression of Red proteins is critical for minimizing the toxic effects of Gam expression...

...networks in biological systems. NGS will also play significantly into the realm of clinical therapeutic...validation process, also known as CRISPR genotyping, is critical to demonstrating causal relationships between ... forms of off-target analysis in order to meet clinical approval. It is key to maintain a consistent set...the detection of KRAS mutations: diagnostic and clinical implications." The Journal of Molecular Diagnostics...

... is necessary for bacterial transformation of chemically and electrocompetent cells is minimal (picogram...nucleic acids (genomic DNA and RNA), proteins and chemical contaminants that have been used during the purification... preparations for this technique must be larger scale, to allow the extraction of enough plasmid to last...addition to being free of any nucleic acid and chemical contaminants, these DNA samples must also be entirely...

...in Plasmids 101, tags can help you determine localization of a protein of interest, purify it, or determine...major caveat - a tag may interfere with protein localization and/or function, so each tagged protein must...proteins found that 20% of tagged proteins did not localize similarly to their untagged counterparts. These...fluorescent-protein tagging show high correlation for protein localization in mammalian cells." Nature methods 10.4 (2013...