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We narrowed to 736 results for: RAN-1

Showing: 341 - 360 of 736 results

  1. Progress Towards a PAM-Free CRISPR

    Type
    Blog Post
    ...nucleotides are represented equally, that’s only a 1/16 chance of having a PAM at a given site. While it... NYN/NRN NYN is less effective than NRN Table 1 - Summary of PAM-flexible Cas enzymes Are we truly...editable with CRISPR technology.     Fig. 1: SpCas9 has sparse PAM coverage in the human genome.... The activity of SpRY was validated in double stranded cleavage and base editing applications, with high...solution? Almost! Assuming a PAM of NRN, 50% of randomly selected genomic loci will have a PAM located ...

  2. Stem Cell Models for Disease & Open Science: Interview with Darrell Kotton

    Type
    Blog Post
    ...Listen to the interview with Darrell Kotton Chapter 1: Introduction to Darrell Kotton and the Boston University...Unit, the Pulmonary Consultation Service, the Alpha-1 Center, and the Pulmonary outpatient clinic, all at...Chapter 3: How all basic research ends up being translational. Ex: the discovery of GFP. Chapter 4: Goals ...

  4. Plasmids 101: Colony PCR

    Type
    Blog Post
    ... for less money. The key steps to colony PCR are: 1) design primers to detect the presence of your insert...primers. There are 3 strategies for primer design: 1) insert-specific primers, 2) backbone-specific primers...previously described. When using insert-specific primers (1), positive clones (+) will give a band, while a negative...control. A good positive control is bacteria transformed with the same backbone plasmid. If this control...strain. A good negative control strain is an untransformed culture of the same strain of bacteria you used...Cloning Technique Learn to Perform a Bacterial Transformation Deposit Your Positive Clones with Us ...

  5. Beyond the Blood Brain Barrier: PHP.B Enters the Central Nervous System Via the LY6A Receptor

    Type
    Blog Post
    ...determined LY6A, also known as stem cell antigen-1 (SCA-1), is the receptor used by PHP.B to reach the CNS...William L. Stanford. "Concise review: stem cell antigen‐1: expression, function, and enigma." Stem cells 25.6...PHP.B has a >40-fold increase in CNS transduction and transduces the majority of astrocytes and neurons... mice whose CNS was transduced by PHP.B but absent in mice who were not transduced by PHP.B. The Deverman...These results were surprising because in vivo transduction of AAV serotypes generally tracks between strains...diverse library of AAV9 mutants that had a small randomized peptide loop inserted in the AAV9 capsid. This... an enhanced version of PHP.B, shows similar transduction efficiency, but at a log lower dose than PHP.B...

  6. Tips for Screening with Yeast Two Hybrid Systems

    Type
    Blog Post
    ...individual activities (1). Researchers also discovered the modular format of some transcriptional activators: that...requirement for transmembrane proteins, a split ubiquitin system has been devised (8). References 1. Casadaban...systems could be combined effectively (3). In Figure 1 for instance, the DBD of the fusion protein shown ...flurry of discovery in the field of eukaryotic transcriptional activation and cell biology. During this period... that the DNA binding domain (DBD) and transcriptional activation (TA) domains retain their individual... regardless of whether it interacts with a transcriptional activator (1a) or a repressor (1b). Scientists...interactions. In later years, improvements to yeast transformation protocols also helped establish the yeast two-hybrid...

  7. Don’t FRET: Bimolecular Fluorescence Complementation Makes Visualizing Protein-Protein Interactions Easy

    Type
    Blog Post
    ...tethered to bring them into close proximity (Figure 1). Formation of the complete FP results in a fluorescent...detected by fluorescence microscopy. Figure 1: How BiFC works. A split FP (e.g., yellow fluorescent... protein interactions in living cells. Nat Protoc 1:1278–1286. https://doi.org/10.1038/nprot.2006.201 ...Forster Resonance Energy Transfer (FRET). Through the non-radiative transfer for energy between neighboring...Whether BiFC constructs are expressed transiently by transfection or stably impacts the success of BiFC..., kinase signaling, integrin signaling, and transcription factor interactions (Kerppola, 2006). One of...interactions between basic leucine zipper and Rel transcription factors (Hu et al., 2002). This would be more...

  8. Light Sheet Fluorescence Microscopy

    Type
    Blog Post
    ...microscopic resolution for studying colloidal gold (1).  The method was based on using a thin plane (sheet...Siedentopf, and R,. Zsigmondy.  (1902) Ann. Phys.  315, 1 – 39.
 A. H. Voie, D. H. Burns, F. A. Spelman. (1993...microscopy/en_de/products/imaging-systems/lightsheet-z-1.html ...microscopy coupled with techniques that make tissues transparent are enabling microscopes to visualize the cellular...the cortico spinal tract, CST, spinal cord and cranial nerve afferents labelled with a AAV8 expressing...sectioning (OPFOS) (2). Arne Voie, David Burns and Francis Spelman focused a laser beam into a thin sheet ...of clear specimens and for in vivo imaging of transparent organisms. Advances in tissue clearing Similar...

  9. Production of Virus in Insect Versus Mammalian Cells

    Type
    Blog Post
    ...centrifugation (lenti) Centrifugation Titration time ~1 day (AAV); ~1 week (lenti) 2–10 days Initial titers Higher...will help you decide which is right for you. Table 1: Mammalian versus insect cells for viral production...only have to worry about cloning your transfer plasmid. The transfer plasmids have different packaging capacities...AAV and lentivirus requires three plasmids — a transfer plasmid containing your gene of interest plus ...in mammalian cells is relatively quick. After transfection, HEK-293T cells start producing virus within...systems, it is standard to do only one passage (transfection). This differs from typical insect cell preparations...an additional couple of days, as it requires transduction of more cells, while AAV can be titrated following...

  10. Hot Plasmids: Summer 2025

    Type
    Blog Post
    ... high brightness and fast maturation time (Figure 1). See the paper for more details on small differences...mGold2t depending on the application. Figure 1: Super-resolution imaging of actin filaments with ...proteins for extended imaging. Nature Communications, 16(1), 3241. https://doi.org/10.1038/s41467-025-58223-5...non-selective anterograde transport to a lack of genetic control. ATLAS (anterograde transsynaptic label based on...of the color spectrum more powerful, the lab of Francois St-Pierre recently developed a pair of new yellow...in other plant species, thanks to the broad host range of TRV. Find TRV-TnpB plasmids here! Weiss, T.,...Viral delivery of an RNA-guided genome editor for transgene-free germline editing in Arabidopsis. Nature Plants...

  11. Designing Your Chalk Talk for the Academic Job Interview

    Type
    Blog Post
    ...acceptable. Aims should be mechanistic, doable by 1-3 people in 1-5 years, and contribute to resolving the overarching...possible, do at least two full length practices (~1 h) and get feedback. It’s also helpful to practice...aims do not need to be grant aims. Each aim could be the basis of a single grant. Either way, think realistically...talk could be your plans for your first NIH R01 grant proposal or a 10-year research plan. I suggest preparing...is a significant need for healthy livers for transplants. Growth of new livers in culture could resolve..., neither of these can substitute for a liver transplant. My lab will combine bioengineered scaffolds ... for presenting a science talk Check out our transferable skills guide Read all of our career blog posts...

  12. Avoiding the Dark Side of Fluorescent Protein Fusions with mOX FPs

    Type
    Blog Post
    ...experiments or even negatively impact cells (see Figure 1). When behaving properly, FPs enable researchers ... disulfide bonds in the secretory pathway (Figure 1) [4,6–8]. Additionally, FPs that encode N-glycosylation... sarcoma-associated herpesvirus.       References 1. Snapp, Erik. "Design and use of fluorescent fusion...excellent choice for fusions with integral membrane proteins and membrane associated proteins (i.e. GPI-anchored...these compartments undergo significant post-translational modifications including glycosylation, disulfide... addition of N-glycans [9]. Both of these posttranslational modifications can disrupt FP folding rendering...including the secretory pathway (Figure 2), the inner membrane space of mitochondria and chloroplasts, the extracellular...

  13. Sharing Your Lab Protocols: Using Apps to Save Time & Track Your Experiments

    Type
    Blog Post
    ...Recently I learned that Addgene’s pLKO.1 cloning protocol is viewed around 3,000 times each month. I ...on the Addgene website in December 2006, the pLKO.1-TRC cloning vector (deposited by David Root of the...would the thousands of other scientists using pLKO.1 variants learn about those updates? This is a problem...restriction digest of plasmid DNA, and bacterial transformation. And more plasmid-related educational resources...

  14. Transferable Skills Guide: Conflict Resolution

    Type
    Blog Post
    ...’re not one of these people yourself.  Fig 1: Quote from Michael Ende, courtesy of morefamousquotes.com... blog Management for ScientistsTransferable Skills Guide: LeadershipTransferable Skills Guide: Teamwork...the right time. Like many of the posts in our Transferable Skills series, this post is not meant to be ... that you could strengthen. The right set of transferable skills, coupled with your technical skills, ... can help, and if and when you find yourself transferring this skill to another job, it will be important...also aggravate long-term personal conflicts.  Transitioning a conflict into a collaboration may not be the...willing and able to solve, don’t take that skill for granted. When you go on interviews or are looking for a...

  15. Better Dyeing Through Chemistry & Small Molecule Fluorophores

    Type
    Blog Post
    ...environments such as live cells and tissues (Figure 1). These flexible strategies give you the best of both...majority of in-cell labeling strategies have two parts: (1) a genetically encoded “tag” expressed as a fusion...Follow him on twitter @rhodamine110.   References 1. Liu, Zhe, Luke D. Lavis, and Eric Betzig. "Imaging...requires more photons. For example, moving from transient overexpression of protein fusions to gene-edited... lipoic acid ligase, biotin ligase, phosphopantetheinyltransferase) – These enzymes catalyze the covalent... tag (Figure 1c). Click chemistry (e.g., transcyclooctene–tetrazine)– Nonnatural amino acids can be ...improved brightness, photostability, and spectral range. Nevertheless, these fluorophores were designed ...

  16. Five Questions to Ask If You're Deciding Between a Master's or PhD Program

    Type
    Blog Post
    ...my master’s, I worked in an academic lab where I ran a handful of assays over and over. Everyday was the...help you decide if a master’s might be for you. 1. Are you interested in master’s-level jobs? Ultimately...your work at conferences, help with manuscript and grant preparation, and lead your own research project....

  17. 27 Hot Plasmids from 2016

    Type
    Blog Post
    ...a rate of up to 1 per 500-1000 bp. In comparison, the DNA replication error rate is 1 per 109 bp. Unlike... and Screening via Transcriptional Output-TANGO (PRESTO-TANGO) to screen the NCC-1 library of approved...(MAGE) (1). MAGE uses recombineering (2) to simultaneously incorporate multiple single-strand DNA (ssDNA...generated 5 common 30bp overlap sequences (Sites #1-5). Each overlap site encodes a unique restriction...First, GMAP-compatible backbones for lentivirus LV 1-5 and retrovirus RV 2-5 were constructed. The authors...of GFP (SEP) by a 9 amino-acid linker (see Figure 1). The key to the biosensor lies in pH: DsRed is relatively...inactivate SEP and leave only DsRed to fluoresce (1). With variants that can be targeted to specific cellular...

  18. What's the Best Way to Elute and Store Your Plasmid DNA?

    Type
    Blog Post
    .... pH and DNases affect DNA stability Figure 1: So many choices! What to do? The main factors ...your downstream experiment. TE (10 mM Tris-HCl,1 mM EDTA, pH 8.0) buffer is the best buffer for preserving...from the DNA. These apurinic sites if in double stranded DNA are repaired by base excision repair, but ...but can lead to mutations in single stranded DNA undergoing replication.   DNase on the other hand can digest...

  19. A New Optogenetic Tool Based on AraC Controls Gene Expression with Blue Light

    Type
    Blog Post
    ... Figure 1: With blue light, the VVD domains of the BLADE dimerize and activates transcription. The ... that dimerizes upon exposure to blue light (Fig. 1). Another blue light-inducible dimerization domain...from one that represses transcription to one that activates transcription.However, this system lacks spatio-temporal...barbara_synbio) May 13, 2021 The original AraC transcriptional regulator Their system is based on the AraC...AraC protein that controls transcription from the PBAD promoter. “AraC is widely used in synthetic biology...It relies on the sugar arabinose to activate transcription by causing conformation changes in the AraC ...your gene of interest, all you have to do is co-transform pBLADEONLY. That way, a previously cloned gene...

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

    Type
    Blog Post
    ... genetic tools in Drosophila (Fig. 1):  GAL4-UAS, a transcription factor binding site. GAL4 expressed...Rodríguez et al., 2011).  Genetics of SPARC Figure 1: SPARC and SPARC2 use the GAL4-UAS system along with...visualize individual/non-overlapping cells. While transgenic animals are commonly used to specifically manipulate... to trigger recombination and expression of a transgene (Lin et al., 2018). However, this system requires...site to cleave and inactivate any read-through transcripts of the effector gene. Effector genes, any gene...across cell types and organisms by replacing the transcription factor binding site and effector in the construct... the SPARC system to use other experimental transcription activation systems, such as LexA/p65 or mCD8...
Showing: 341 - 360 of 736 results