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We narrowed to 1,010 results for: CAL;

Showing: 541 - 560 of 1010 results

  1. To Codon Optimize or Not: That is the Question

    Type
    Blog Post
    ...expression limiting regulatory elements or use a non-canonical code (Gustafsson et al., 2004), such as Hoogsten...recognize multiple synonymous codons) may actually be critical for determining the rate of translation. The ribosome...10.1101/gr.139758.112 Mauro VP, Chappell SA (2014) A critical analysis of codon optimization in human therapeutics... GluCl ion channel genes for mammalian cells dramatically improves expression levels. Journal of Neuroscience...synthetic? How Addgene;s dataset reveals trends in biological innovation Resources on Addgene.org Check out...

  2. Bioinformatics at Addgene

    Type
    Blog Post
    ...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 ...

  3. New CRISPR Tools: Cas7-11 and PASTE

    Type
    Blog Post
    ...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...

  4. Deciphering the Mysteries of Behavior with Viral Vectors

    Type
    Blog Post
    ...regions and allocate specific functions to them. Electrical recordings and lesioning studies have been essential...within the LC of mice. When those neurons are chemogenetically activated, norepinephrine release causes a...an enhanced propensity for movement and activity called locomotor sensitization. This enhanced movement...responsible for the antidepressant effect. To specifically activate the Drd1 neurons, they injected AAV2... "Hypothalamic CNTF volume transmission shapes cortical noradrenergic excitability upon acute stress."...

  5. Plasmids 101: Shuttle Vectors

    Type
    Blog Post
    ...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...

  6. CRISPR 101: Any Base Transversion Editors

    Type
    Blog Post
    ...product purity of 66%. Pro tip! Abasic sites are also called apurinic or apyrimidinic sites and abbreviated ...are the same protein!) (Chen et al., 2024). They called their initial editor AXBE, for A → X base editor...in a G → Y (Y = T or C) base editor, which they called glycosylase-based guanine base editor (gGBE) (Tong... Therapeutics team has deposited their editors, called gTBE and gCBE, with Addgene. gTBE can introduce...site A DNA site with the nucleobase removed. Also called an apurinic / apyrimidinic (AP) site. UNG / UDG...

  7. CRISPR 101: Targeting RNA with Cas13a (C2c2)

    Type
    Blog Post
    ...post was updated on Jul 27, 2020. CRISPR, and specifically Cas9 from S. pyogenes (SpCas9), is truly an ...specified by the crRNA sequence, it adopts an enzymatically “active” state rather than reverting to an inactive...propensity of Cas13a to bind and cleave RNA non-specifically. The ability of dCas13a to bind target RNA without...modulating splicing, and regulating expression of specifically targeted RNAs. Finally (and perhaps most interesting...observed shRNA off-targets. They subsequently used catalytically inactive dLwaCas13a as a programmable RNA-binding...

  8. Tips for Using FRET in Your Experiments

    Type
    Blog Post
    ...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. ...

  9. SARS-CoV-2/COVID-19 Detection Methods Based on CRISPR/Cas

    Type
    Blog Post
    ... 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...

  10. Transferable Skills Guide: Conflict Resolution

    Type
    Blog Post
    ...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...

  11. Evolution of Brainbow: Using Cre-lox for Multicolor Labeling of Neurons

    Type
    Blog Post
    ...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...

  12. Viral Vectors 101: AAV Serotypes and Tissue Tropism

    Type
    Blog Post
    ...target specific tissues, even when administered systemically, without causing disease? Recombinant adeno-...genomic structure of AAV2, so they are formally called rAAV2/1, rAAV2/2, and so on, although we often ...in-stock viral prep service.  AAV5 AAV5 is the most genetically distinct AAV. Though AAV5 is one of many serotypes...on the AAV capsid. Molecular Therapy. Methods & Clinical Development, 26, 331–342. https://doi.org/10.1016...is required for gene transfer. The Journal of Biological Chemistry, 276(23), 20610–20616. https://doi....

  13. Generating Mouse Models Using CRISPR/Cas9

    Type
    Blog Post
    ...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...

  14. Addgene Protocols Go Viral: AAV Purification and Titration

    Type
    Blog Post
    ...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...

  15. Tips for Working in Industry & Success in Management: Interview with Crystal Shih

    Type
    Blog Post
    ...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...

  16. Alex Chavez on MD PhDs, CRISPR, Gene Drives, & More

    Type
    Blog Post
    ...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...

  17. Lambda Red: A Homologous Recombination-based Technique for Genetic Engineering

    Type
    Blog Post
    ...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...

  18. Sequencing Options for CRISPR Genotyping

    Type
    Blog Post
    ...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...

  19. Plasmid Preps: Different Purity, Different Quantities, Different Uses

    Type
    Blog Post
    ... 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...

  20. Inntags: Innovative Protein Epitope Tagging

    Type
    Blog Post
    ...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...
Showing: 541 - 560 of 1010 results