We narrowed to 356 results for: SAM activators
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TypeBlog Post...expressed, LacZ𝛼 complexes with LacZ𝜔 and forms an active LacZ tetramer.  This tetramer cleaves chlorophenol...evolution to create APEX2. They have now applied the same directed evolution technique to the biotin ligase...
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CrispyCrunch: High-throughput Design and Analysis of CRISPR+HDR Experiments
TypeBlog Post...HDR may inadvertently re-create a match for the active guide. With these constraints in mind, we implemented...scientists design and analyze batches of CRISPR samples. We invite you to jump in and try it out, or take... databases, including primer3, biopython, bwa, samtools, bowtie, UCSC genome browser, Ensembl, and others...currently Crispor and Crispresso, that adhere to the same principles listed here. It employs biological best...productivity in the Biohub. We hope it will do the same for you. Send us your questions or comments to gdingle... -
Story of a SynBio Startup: RevBio's Epiphany (or Lack Thereof)
TypeBlog Post...commission in the United States Air Force. She left active duty to pursue a degree in a synthetic biology ... synthetic biology lab Keira and I worked in the same lab for four years. It was a cutting-edge synthetic... on the detector plants, we expected to see that same anger. Instead, social media comments were more ... -
Antibodies 101: Conjugation
TypeBlog Post...each of these enzymes allow you to detect enzyme activity (and therefore your antibody) either through a... antibody will have been verified to perform the same as its unconjugated version, but if you find yourself... -
Mouse Modeling, Part 1: Genetically Engineered Mice
TypeBlog Post... include tetracycline or doxycycline mediated activation of Cre to either induce or repress Cre expression...repair is a random process, zygotes edited using the same nuclease may not result in genetically identical... -
Fluorescent Tagging of Endogenous Genes with SapTrap
TypeBlog Post...Beyond simply monitoring protein localization and activity, CRISPR tagging systems can be modified to assay... 2-3). Since donor plasmids of the same type will produce the same unique SapI 5’ overhang, different ...about Fluorescent Protein Timers Check out Photoactivatable Fluorescent Proteins Use Rosella to Monitor... -
Adenoviral Delivery of CRISPR/Cas9 Aims to Expand Genome Editing to Primary Cells
TypeBlog Post... Doench CRISPR-Cas9: Tips for Optimizing sgRNA Activity by John Doench All CRISPR Posts  ...Beatty et al., 2012). These features make AdV an attractive candidate for in vivo delivery of advanced genome...achieved by isogenic AdVs encoding TALENs targeting the same chromosomal region. The CRISPR/Cas9-derived RNA-...in a second paper published in Nature Methods the same year that delivering RNA-guided nucleases or TALENs... -
Your Lentiviral Plasmid FAQs Answered
TypeBlog Post...proteins that target specific tissues are areas of active study (for a review click here). Alternative envelopes...often: Q1: Are retroviruses and lentiviruses the same thing? Can I package lentivirus with retroviral ...both lentiviruses and gamma-retroviruses use the same genes for packaging (that's gag, pol, and env), ...initial infection. SIN is shorthand for self-inactivating, which is achieved by deleting a large portion... -
Viral Vectors 101: The Retroviral Lifecycle
TypeBlog Post...lines in which each cell has the capacity to be activated to produce infectious murine leukemia virus. Virology...instructions in the same producer cells as the virus. Because it is made at the same time as the other ... a retroviral particle, which is essentially the same for viruses and viral vectors. Key to the wild-type... -
Negative Can Be Positive: Open AAV Data with Addgene
TypeBlog Post... one’s work. As a community, we must therefore actively work to remove the stigma around sharing negative...well for a neighboring cell type, even within the same brain region or nucleus. Just optimizing AAV expression...vectors, many from people trying to answer the exact same question I was: What AAV vector is best for my specific...wanted to see more negative data (Figure 2.) At the same time, when scientists who support open science, ... -
Negotiating Work and Life: How to Find the Joy
TypeBlog Post...one area at a time and try to say “no” to other activities. Take care of yourself. Take a vacation alone...whatever works for you, but most days go home at the same time. Use time management tools and tactics to plan...need to match up exactly when you fold them. The same with sheets – wash them and put them right back ...family by forming a social circle of people in the same situation. Go to church, join a club, meet the neighbors... -
Using AAV for Neuronal Tracing
TypeBlog Post... reciprocal, forming a neuronal loop which is activated and strengthened during memory formation and memory...connections that are functionally relevant, i.e. activity-dependent neuronal tracing. Apart from mapping...al., 2008) AAV6 (Towne et al., 2010), AAV8, -9 (Masamizu et al., 2011); AAV5 (Aschauer et al., 2013). Anterograde...PMCID:Â PMC5529245. Choi, V.W., McCarty, D.M., and Samulski, R.J. (2005). AAV Hybrid Serotypes: Improved Vectors...PMC2895694. Hollis, E. R. 2nd, Kadoya, K., Hirsch, M., Samulski, R. J., and Tuszynski, M. H. (2008). Efficient...PMID:Â 25374356. PubMed Central PMCID:Â PMC4269290. Masamizu, Y., Okada, T., Kawasaki, K., Ishibashi, H., Yuasa...PubMed Central PMCID:Â PMC5572074. Salegio, E.A., Samaranch, L., Kells, A.P., Mittermeyer, G., San Sebastian... -
Adeno Associated Virus (AAV) for Cell and Gene Therapy
TypeBlog Post...host chromosomes, further minimizing potential activation of innate immunity. AAV vectors are also capable... PubMed Central PMCID: PMC3152950. 3. Jacobson, Samuel G., et al. "Improvement and decline in vision with... -
Overcoming the Challenges of Lentiviral Production
TypeBlog Post...system incorporates the Tet-off transcriptional activation system to drive high levels of expression of ... 10 cm2 dishes. While this option is attractive because the same procedure for a single 10 cm2 dish can...systems such that it no longer requires the viral transactivator, Tat. For more information on the different... -
The AAVantages of AAV in CRISPR Screens
TypeBlog Post...wide variety of screens, including interference, activation, and knockout. Most CRISPR screens rely on ...expression of lentiviral vectors make them an attractive option. However, safety requirements and expression...vivo. The general process of these screens is the same: Deliver CRISPR library to cells of interest. Select...Low, S. H., François, A., Suárez-Amarán, L., & Samulski, R. J. (2022). AAV vectors: The Rubik’s cube of... -
Plasmids 101: Secondary Nanobody Toolbox
TypeBlog Post...State University Use Nanobodies to Control Protein Activity Learn more about protein tags Read about E. coli...figures 4A-C). And since they have limited cross-reactivity between different IgG subclasses and different...primary antibody-nanobody mix can be added to your sample, thus eliminating the secondary antibody incubation...568, or Alexa Fluor 647 before application to the sample. Staining with this multi-color staining workflow... -
AAVs in Retinal Gene Therapy
TypeBlog Post...back, better and safer. It is now one of the most active and exciting areas of science. This is hopefully...retinal cells - AAV - were becoming available. At the same time advances in human genetics led to the identification...24664759. PubMed Central PMCID:Â PMC4267879. 10. Samulski RJ, Muzyczka N., 2014. AAV-Mediated Gene Therapy... -
Tips for Using FRET in Your Experiments
TypeBlog Post...used to measure conformational changes, cleavage activity using FRET-based biosensors, and interactions ...What is FRET? In their JCB 2003 paper, Sekar and Periasamy defined FRET as "a distance-dependent physical...341-50. doi: 10.1002/bies.201100098. Sekar RB, Periasamy A. Fluorescence resonance energy transfer (FRET... -
Empty Backbones - Choosing Your Perfect Plasmid Backbone
TypeCollection... reporter plasmid with very low basal activity shRNA/RNAi Gene silencing Browse RNAi empty vectors... Mammalian pLX304 - 3rd gen lentiviral vector lentiSAMv2 - Lentiviral sgRNA cloning backbone pLKO.1-blast... -
Antibodies 101: Reading a Flow Plot
TypeBlog Post... dendritic cell function and increased B cell activation. PLOS ONE, 12, e0180664. https://doi.org/10.1371... are represented as a series of dot plots in the same order. However, in an actual flow cytometry run,...parallel. Let’s imagine we’re looking for cells in a sample that are negative for marker A and positive for... proxy for nucleic complexity of the cell. The sample will have been labeled using a panel of markers... dots. In a contour plot, each line contains the same number of events. Thus, contour lines that are close...