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We narrowed to 919 results for: IRE;

Showing: 441 - 480 of 919 results

  1. Designing Your Chalk Talk for the Academic Job Interview

    Type
    Blog Post
    ...This post was contributed by Erik Snapp, the Director of Student and Postdoctoral Programs at the Janelia...single model. The first approach can lead to new directions, while the second approach could terminate with...think realistically about the time and resources required to feasibly accomplish the aims and clarify what... and Structural Biology, and now serves as the Director of Student and Postdoctoral Programs at the Janelia...

  2. Rett Syndrome: A History of Research and Therapeutic Outlooks

    Type
    Blog Post
    ...Rett syndrome experience regression of previously acquired skills such as speech, coordination, purposeful...programs in biopharma. AveXis, co-founded by Kaspar, acquired by Novartis and rebranded to Novartis Gene Therapies...adeno-associated viral (AAV) vector. The editase repaired half of the mutant MECP2 RNA in each of three ...Samaco RC, Zoghbi HY (2011) Adult Neural Function Requires MeCP2. Science 333:186–186 . https://doi.org/10.1126...

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

    Type
    Blog Post
    ... the ideal codon for every amino acid across an entire polypeptide. Many researchers have created codon-optimization...necessary for proper protein folding (Stadler and Fire, 2011). Therefore, codon optimization may disrupt...1987) The codon adaptation index-a measure of directional synonymous codon usage bias, and its potential...doi.org/10.1016/s0165-0270(02)00362-x Stadler M, Fire A (2011) Wobble base-pairing slows in vivo translation...

  4. Four Base Editing Reporters to Monitor and Enrich Editing in Real-time

    Type
    Blog Post
    ...DNA strain which results in two DSBs that can be repaired by NHEJ restoring mCherry expression. Cells can...populations. To develop this method the lab was inspired by work that used an integrated GFP reporter that...fluorescent protein) upon CRISPR/Cas9 homology directed repair (HDR) (Glasser et al., 2016). For TREE ...different reporter proteins. Protein translation requires the start codon of AUG immediately downstream ...

  5. Transferable Skills Guide: Creativity

    Type
    Blog Post
    ...skill developed during PhD training. Creativity is required to develop new projects and test novel approaches... all the funding in the world? What if you could hire more people? What would you do with unlimited time...over half of which, when implemented, impacted the entire company. It is important to note that the person...product team is an incredibly innovative group. Our entire company benefits from their policy! Perfectionism...

  6. Harnessing TnpB for Genome Editing: A Compact and Versatile Tool for Your Research

    Type
    Blog Post
    ...cleave both strands. Similar to how Cas proteins require a PAM (protospacer adjacent motif) near the target...a different TAM (5′-TTAT-3′). The unique TAM requirements expand the number of genomic sites that can ... and prime editing (Molla et al., 2021), which require fusing Cas proteins with additional enzymes or ...transformation into plant cells. Conclusion & Future Directions TnpB represents a new generation of compact, ...

  7. QC Sequencing Technologies at Addgene

    Type
    Blog Post
    ...strand is synthesized (Figure 1). Sanger sequencing requires four separate reactions, where a fraction of one...in ultra-long read mode), enough to sequence an entire plasmid in one read. Runs can be completed in anywhere...sequencing, as well as the ability to sequence an entire viral genome in one read. We also use Nanopore ...keeping up with the large amount of QC analysis required every week. They are constantly exploring ways...

  8. Plasmids 101: Shuttle Vectors

    Type
    Blog Post
    ...helps increase cellular uptake, but this approach requires compatible cell lines that express viral machinery...ribosome. One resistance gene on your vector might require two different selection molecules! It all depends...option is to use a different method of selection entirely. Auxotrophic selection relies on the host organism...organism being unable to synthesize a nutrient required for growth. Before delivery of the vector, the deficient...

  9. Plasmids 101: Plasmid Incompatibility

    Type
    Blog Post
    ...  Antisense RNAs encoded on the plasmid either indirectly inhibit replication through inhibiting translation...translation of replication machinery proteins, or directly inhibit replication by binding to the origin of...sequences within the origin of replication and are required for replication in plasmids that contain them ...Kamruzzaman M, Shoma S, Thomas CM, Partridge SR, Iredell JR (2017) Plasmid interference for curing antibiotic...

  10. Four Ways to Package Transgenes That Exceed the Size Limit of Adeno-associated Virus

    Type
    Blog Post
    ...AAV donor template for CRISPR-mediated homology directed repairs (HDR). Split AAV vector approaches Split...trans-splicing (62%) vector. 4. Sequential homology directed repairs Overview: The sequential homologous recombination... donor templates using two sequential homology directed repairs (HDR) with CRISPR/Cas9. Donor A contains...M, Colella P, Garcia-Hoyos M, Gibbs D, Kim SR, Maguire A, Rex TS, Di Vicino U, Cutillo L, Sparrow JR, ...

  11. Hot Biosensors 2022: Year-End Roundup

    Type
    Blog Post
    ...variety of behaviors, opening up new research directions that were inaccessible with earlier tools. They... in localization to the cell membrane and not a direct change in the fluorescent properties, pYtags are...variations and heat transfer at the subcellular level requires a probe with fast kinetics and high signal/noise...temperature indicator B-gTEMP designed to meet those requirements (Lu et al. 2022). By fusing two bright fluorescent...

  12. Plasmids 101: Expressing shRNAs for RNA Interference

    Type
    Blog Post
    ...composition of the RISC varies, but at the very minimum requires an Argonaute protein. This Argonaute protein is...shRNAs using plasmids The design of shRNA plasmids requires three important steps: Designing the shRNA sequence...This is much shorter than miRNA plasmids, which require additional flanking regions comprised of upstream...35(9), 872–878. https://doi.org/10.1038/nbt.3947 Fire, A., Xu, S., Montgomery, M. K., Kostas, S. A., Driver...

  13. Viral Vectors 101: Integrase-Deficient Lentiviral Vectors

    Type
    Blog Post
    ...Ciuffi, 2012). One family of viruses, retroviridae, require integration for their life cycle to continue. They...IN (integrase). These viral packaging genes are required for the production of lentiviral particles.  The... keep in mind is that titering these IDLVs may require a slightly different protocol than regular lentiviral... (2008). Insertional mutagenesis combined with acquired somatic mutations causes leukemogenesis following...

  14. Antibodies 101: Single Chain Fragment Variables (scFvs)

    Type
    Blog Post
    ...be expressed in bacteria, while mAbs generally require mammalian expression systems. scFvs are also small...to avoid animal use by using semi-synthetic or entirely synthetic VL and VH sequences. Phage display also...for the scFv (Gaciarz and Ruddock., 2017), but requires an oxidizing environment to form. When expressed... age-related macular degeneration (AMD) since direction injection to the eye results in high drug concentrations...

  15. Multiple Plasmids at a Low Price: Kits from Addgene

    Type
    Blog Post
    ...bypasses the 5′ mRNA cap required for eukaryotic systems and instead directly engages the ribosome for... protocol and are compatible with single-end or paired-end read flow cells. FREQ-Seq is an open source... The kit contains all the variant PUF segments required to bind any 8 base RNA sequence. The segments ...

  16. Antibodies 101: Multiplex Immunofluorescence

    Type
    Blog Post
    ...secondary antibodies altogether and multiplex with direct IF, using primary antibodies that are conjugated...However, the primary-secondary workflow (a.k.a. indirect IF) is more flexible and efficient, since you ...primaries for whichever targets you like. Plus, indirect IF amplifies the signal since multiple secondary...design. Like selecting an antibody, some of the requirements are similar to standard IF. Here, we’ll walk...

  17. Plasmids 101: Dimers and Multimers

    Type
    Blog Post
    ... on as a dimer, depicted on the right, with the entire monomeric sequence duplicated in tandem, resulting... with large inserts (>7 kb)(Berg et al., 1989), direct repeats (Ribeiro et al., 2009), and at high copy...frequency plasmid recombination mediated by 28 bp direct repeats. Mol Biotechnol. 2008 Nov;40(3):252-60....Summers D. Timing, self-control and a sense of direction are the secrets of multicopy plasmid stability...

  18. Viral Vectors 101: AAV Serotypes and Tissue Tropism

    Type
    Blog Post
    ...proteoglycan (HSPG)3 AAV receptor (AAVR; not strictly required)2 Human fibroblast growth factor receptor ...targeting of projection neurons and enhances astrocyte-directed transduction. Nature Communications, 14(1), 3792...Adeno-Associated Virus Serotype 4 (AAV4) and AAV5 Both Require Sialic Acid Binding for Hemagglutination and Efficient...adeno-associated virus type 5 to 2,3-linked sialic acid is required for gene transfer. The Journal of Biological Chemistry...

  19. Antibodies 101: The Basics of Western Blotting

    Type
    Blog Post
    ...overnight.  In contrast, a semi-dry transfer only requires enough buffer to saturate the sandwich and typically...without any transfer buffer. These systems, however, require you to purchase expensive system-specific sandwiches...on the antibodies and labels being used and may require some trial and error (Yang et al., 2012).  Choosing... they provide qualitative measurements. If you require precise quantification of a protein, consider using...

  20. Finding Your Perfect Job After University

    Type
    Blog Post
    ... degree. I loved parasitology. However, when I enquired about roles in this field, most jobs needed at...understood why parasitology roles in the field require experience. Working in industry: Microbiology ...redundancies can be made within the company; new staff hired in anticipation of the contract suddenly have less...institutes. The lab at the University of Queensland required a scientist for a 6 month project to validate ...

  21. Mycoplasma Contamination: Where Does It Come From and How to Prevent It

    Type
    Blog Post
    ... to stain all DNA. Some amount of expertise is required in this process because result interpretation ...serum to be used for subculturing. Avoid using expired products or reagents that have been stored for ...cross-contamination. Make sure you keep all the required equipment and reagents inside the laminar airflow... Biotechnology.       References Degeling MH, Maguire CA, Bovenberg MSS, Tannous BA (2012) Sensitive ...

  22. Typing CRISPR Systems

    Type
    Blog Post
    ...systems has led to extreme sequence divergence that requires a multi-pronged analysis to sort effectively. ...efficiency when conducting edits using homology-directed repair. There are 10 subtypes of type V, named...select RNA transcripts, or be engineered to conduct direct RNA editing. There are four subtypes, A–D, with...surface-level function of each type could lead you to new directions in your research. We all have our preferences...

  23. SciComm with the Experts at Science in the News Part 1

    Type
    Blog Post
    ...University. Effective science communication is required to promote public support for research and to ...graduate student at Harvard University, former Co-director of Science in the News, and member of the SITN...her PhD from Harvard University, is a former Co-director of Science in the News, and is a member of the...

  24. Expanded Access to Rabies Plasmids

    Type
    Blog Post
    ... rabies materials may be required for some countries and may not be required for others. You will be notified... notified if an export license is not required for your country after checkout, and the processing fee...

  25. Addgene’s Newest Way to Speed Your Research: The Cloning Grade DNA Service

    Type
    Blog Post
    ...after receiving the plasmid from Addgene you could directly start your digest, PCR, or transformation?  Great... we decided to include 10 plasmids that can be directly used in the research on the SARS-CoV-2. Nine of...experiments such as cloning, digestions, PCR etc. and/or directly long term storage. Every preparation is quality...

  26. The Developmental Studies Hybridoma Bank: Over 25 Years of Antibody Sharing

    Type
    Blog Post
    ...The Developmental Studies Hybridoma Bank (DSHB), directed by David R. Soll, is a repository for monoclonal...most popular monoclonal antibodies include those directed against human transcription factors such as PAX7...to Our Guest Blogger!   David Soll has been the Director of the Developmental Studies Hybridoma Bank (DSHB...

  27. Michael Koeris' Journey from Grad Student to Entrepreneur: The Story of Sample 6

    Type
    Blog Post
    ... taking up a position on the Addgene Board of Directors. We're excited to watch both Addgene and Sample...How to Be A Good Manager from Addgene Executive Director Joanne Kamens Resources on the Addgene Website... Check Out the Other Members of our Board of Directors Become a Part of the Addgene Community ...

  28. Science Communication for Everyone

    Type
    Blog Post
    ...every science field as we expand and encourage more direct conversations between scientists and non-scientists...process of starting a forum where non-scientists can directly approach scientists and doctors with their questions.... Use hashtags effectively. This would help you direct your content to the right audience. Avoid using...

  29. Designer PUF Proteins for Any RNA Target

    Type
    Blog Post
    ...meteoric rise of CRISPR technology, the ability to direct enzymes – from nucleases to transcription factors...RNA. As a result, targeting the RNA of interest requires that the RNA be tagged. “This tool makes it possible...She also notes that for reasons that are not yet entirely clear some PUF proteins lose their solubility....

  30. Optogenetics + CRISPR, Using Light to Control Genome Editing

    Type
    Blog Post
    ...) that can be repaired by either non-homologous end joining (NHEJ) or homology directed repair (HDR). ...CIB1 fusion binds to the target DNA sequence as directed by the guide RNA (gRNA), while the CRY2PHR-activator...employed in other split systems where light is required to reconstitute enzymatic activity. Photoactivatable...

  31. Management for Scientists: Seeking Feedback

    Type
    Blog Post
    ...information to make improvements. Members connect directly with one another – not just with the team leader...'t work to get people sharing.   Frequently ask direct questions for specific feedback in non-public settings...Scientists. Subscribe to the Addgene Blog to follow the entire series as it is published. For more reading try...

  32. Celebrating Accomplishments in the Lab

    Type
    Blog Post
    ...work, determination, passion, and patience are required to build and lead a productive and successful ...sharing meals, going on trips, and going to New Hampshire in the summer for the annual lab retreat. How ...no play Although hard work and perseverance are required to achieve success in the lab, it is important...

  33. IBEX Knowledge-Base: A Data Resource for Multiplex Tissue Imaging

    Type
    Blog Post
    ...to dive into conversations with other users. The entire dataset can be downloaded from Zenodo, including... the Reagent Resources table or by downloading directly from the Zenodo dataset.     Figure 2... and answer questions. Andrea and Ziv were inspired by the open-software community when designing ...

  34. Using Ultrasound to Image Bacteria in vivo: Acoustic Reporter Genes

    Type
    Blog Post
    ... in the body, like the gastrointestinal tract. Require Administration of Substrate: Some luciferase reporters... such as Renilla luciferase (RLuc) or NanoLuc, require injection of substrate to produce bioluminescence...arg2 vesicles is the amount of soundwave pressure required to collapse them: arg2 vesicles collapse at a ...

  35. Sensing Neuronal Dopamine

    Type
    Blog Post
    ... the main source of the body’s DA. Neurons often fire at a certain frequency, and this frequency can indicate...frequency of its release) are well understood, the direct effect of DA binding to a target neuron is more...the region of the brain where DA quantitation is desired (Figure 1). The probe perfuses liquid, which causes...

  36. Early Career Researcher Toolbox: Free Tools for Making Scientific Graphics

    Type
    Blog Post
    ...are under a Creative Commons 3.0 license, which requires users to give appropriate credit, provide a link...indicate if changes were made to the images. This requirement means the images are probably better suited for... image search is a great tool for searching the entire internet for scientific graphics. The tools settings...

  37. Genetically-encoded Sparse Cell Labeling - A SPARC of Innovation

    Type
    Blog Post
    ...transgene (Lin et al., 2018). However, this system requires precise titration of the AAV. So far, tools to...to overcome this challenge in Drosophila require heat-shocking the system or using chemical inducers of... that will be labeled in a given population is directly correlated to the length of the truncated attP...

  38. Hot Plasmids and Viral Preps - July 2021

    Type
    Blog Post
    ...    Novel fluorescent enzyme biosensors provide direct, specific and sensitive detection of the plant ...strigolactones (SLs) are costly, non quantitative or require heavy optimization. Claudia Vickers’s lab has designed...than on the fluorescence intensity, it provides a direct and quantitative detection of SLs. The authors ...

  39. New CRISPR Web Resources and #12DaysofCRISPR Recap

    Type
    Blog Post
    ... Matchmaker Day 2: Homology directed repair: CRISPR 101: Homology Directed Repair and 3 Tips to Improve...101: Multiplex Expression of gRNAs and The PAM Requirement and Expanding CRISPR Beyond SpCas9 Day 8: Repair...
Showing: 441 - 480 of 919 results