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We narrowed to 736 results for: RAN-1

Showing: 461 - 480 of 736 results

  1. CRISPR 101: Cytosine and Adenine Base Editors

    Type
    Blog Post
    ...other. Figure 1: Base transition editors catalyze purine-to-purine or pyrimidine-to-pyrimidine base...changes require Base Transversion Editors, which we've discussed in two follow-up posts: (1) (2). Base editors...SpCas9 base editors with editing windows as small as 1-2 nucleotides. To reduce off-target effects associated...engineering and protein delivery. Nature Communications, 8(1), 15790. https://doi.org/10.1038/ncomms15790 Richter...the opposite strand). ABEs make an A to G change (or a T to C change on the opposite strand). Both are ...place on the free strand of DNA and converts a C to U without producing a double strand break. UGI = uracil...Repair CRISPR 101: Cytosine Transversion Editors CRISPR 101: Any Base Transversion Editors Learn about base...

  2. Supporting Reproducibility with a Connected ELN

    Type
    Blog Post
    ...true reproducibility. The following are critical: (1) connectivity to other data sources used in research...engagement with the Wisconsin research community were to: (1) prevent vendor lock in by making it easy to get data...that emerged from intensive engagement with a wide range of people at the University of Wisconsin in 2011...reproducibility an ELN should have integrations with a broad range of relevant generic and science-specific tools. ... however, in some cases it would be possible to grant those wanting to understand and/or attempt to reproduce...Massachusetts, and feels privileged to be part of the vibrant life sciences research and commercialization community...

  3. Adenovirus Guide

    Type
    Guide
    ...to an antigen. AdEasier-1 cells BJ5183 E. coli cells containing the pAdEasy-1 packaging plasmid. Adenovirus...enzyme PmeI . This construct is then transformed into AdEasier-1 cells (Addgene #16399) , which are BJ5183...plasmid pAdEasy-1 (Addgene #16400) is suitable for most purposes. For especially long transgenes, the use of...gene therapy landscape . Signal Transduction and Targeted Therapy, 6 (1), 53. https://doi.org/10.1038/s41392...icosahedral capsids and build new virions. Figure 1: Wild-type adenovirus and its genome. Genes over or...production system. Figure is based on the use of pAdEasy-1. Created with BioRender.com. AdMax™ AdMax™ is another...as a Zika virus vaccine . Scientific Reports, 10 (1), 3613. https://doi.org/10.1038/s41598-020-60238-5...

  4. New Viral Vectors - Spring 2025

    Type
    Blog Post
    ... new preps might enhance your research!   Table 1: New ready-to-use viral vectors in the Addgene repository...diverse needs of our community and support a wide range of experimental designs. Here are nine great additions...Request service, which offers over 3,000 eligible transfer plasmids and five different serotypes. Not sure...

  5. Of Myc and Men

    Type
    Blog Post
    ...more applications.   Fig. 1 Making a Protein Tag (Abridged) (1) Locate a promising monoclonal antibody...an amide group added to its C-terminus post-translationally, which turned out to be critical for antibody...

  6. Plasmids 101: Protein tags

    Type
    Blog Post
    ...does not greatly alter the size of the tag.   Table 1: Common protein tags Tag Epitope Mass (kDa) Function... conditions FLAG DYKDDDD or DYKDDDDK or DYKDDDK 1 Affinity and Purification Good for antibody-based ...usually smallish peptides incorporated into a translated protein. As depicted in the accompanying cartoon...peptide-based proteosome inhibitors, glutathione S-transferase (GST), which can be fused to recombinant proteins...

  7. Reaching out to China: Canton Nucleic Acids Forum (CNAF) 2015

    Type
    Blog Post
    ...fragments of these transcripts. I remember when small RNA was just something we ran off the bottom of ...viruses - 2 cas systems regulate at the DNA level and 1 at the RNA level. Udi Qimron from Tel Aviv University...’t give tRNAs much thought. High throughput transcriptome sequencing methods including work done by the...hypothesized that life started with RNA and a simple cell membrane. His recent work shows that they are making fairly...knew…I had barely heard of these). These single stranded, covalently closed RNAs seem to be conserved in...

  8. 28 Hot Plasmid Technologies from 2015

    Type
    Blog Post
    ...dark affinity range (130nM to 4.7µM for iLID nano and 800nM to 47µM for iLID micro; Figure 1). With this... August 20th; 14(1). TRICK: A method for visualizing the first round of translation in living cells ...light-sensitive LOV2 domain of Avena Sativa phototropin 1 (AsLOV2). In the dark, this peptide is caged by the...Figure 2). They are all available now at Addgene. 1 Lungu et al., Chem Biol. 2012 Apr 20;19(4):507-17....Guntas et al., Proc Natl Acad Sci USA 2015 Jan 6; 112(1):112-7. doi: 10.1073/pnas. CRISPR-Cas9 optogenetic...for a variety of experimental purposes including (1) single or multiple, constitutive or inducible, gene...the years. The authors optimized these systems by: 1) decreasing the number of plasmids required for each...

  9. Advanced Uses of Cre-lox and Flp-FRT - A Neuroscientist’s View

    Type
    Blog Post
    ...single neurons in the living brain   Figure 1: Plasmid mix to label neuronal morphology (eYFP) and...genOway press release for more information. References 1. Atasoy, D., Y. Aponte, H. H. Su, and S. M. Sternson...concentration of FLEx vectors guarantees that most cells are transfected with all labeling plasmids and...PMCID: PMC2593125. 2. Buchholz, F, P O Angrand, and A. Francis Stewart. 1998. “Improved Properties of ... sites can lead to excision, inversion, and translocation of DNA in a similar fashion as between loxP ... has a temperature optimum of 37 °C (Buchholz, Angrand, and Stewart 1998). FLPe’s performance was further...in FLPO, which shows similar efficiency to Cre (Kranz et al. 2010). These differences in recombinase efficiency...

  10. Antibodies 101: Reading a Flow Plot

    Type
    Blog Post
    ...Figure 1). Clustered events are called a population.                             Figure 1: While...conceptualize where the populations are, try drawing a quadrant over the plot that divides it into four, using...different populations by imagining (or drawing) a quadrant over the dot plot. Created with biorender.com....Biorender! While a helpful visualization tool, a quadrant will often be too clean for real-world data. Most...and debris. The side scatter roughly measures granularity, a proxy for nucleic complexity of the cell.  ...these populations, I suggest using the imaginary quadrant to help (Figure 5). Even though this real-world...real-world data is far messier than biorender plots, quadrants can still help me as a reader if I'm struggling...

  11. Lentiviral Vector Guide

    Type
    Guide
    ... typically ranges from 8–10 kb, encoded on RNA (Figure 1). This RNA is reverse transcribed into the provirus...— contains transgene and LTRs (chimeric 5’ LTR) Packaging plasmid 1 — contains gag and pol Packaging plasmid...pre-infusion CAR T-cell products. Journal of Translational Medicine , 20 (1). https://doi.org/10.1186/s12967-022...been removed or mutated for increased safety. Figure 1: Wild-type lentivirus genome. Created with BioRender.com...genes (for example, vif , vpr , vpu , and nef for HIV-1). The LTRs flank all of these genes, and anything ...and third-generation systems, please refer to Table 1. Figure 4: Third-generation lentiviral plasmids. Created...heterologous enhancer/promoter such as CMV or RSV Table 1. Summary of key differences between second- and third-generation...

  12. Podcast: Interview with Addgene's Founders

    Type
    Blog Post
    ...Addgene's founders Podcast breakdown Intro Chapter 1 - Who are Addgene's Founders? Chapter 2 - The process...Chapter 3 - Addgene as a leader in standardized tech transfer and the adoption of the UBMTA Chapter 4 - Developing...ensuring that success Chapter 6 - Developing Addgene’s Brand and culture Chapter 7 - Building Addgene as a global...

  13. CRISPR Activation: A Practical Guide

    Type
    Blog Post
    ...deficient in DNA cutting activity to a transcriptional activation domain (Fig 1). Two ways to use CRISPRa for... (2016) Targeted HIV-1 Latency Reversal Using CRISPR/Cas9-Derived Transcriptional Activator Systems. PLoS...promoter of another gene, thereby driving aberrant transcription. That being said, we generally pick guides...been significant work on constructing Cas9 transcriptional activators. These constructs allow for the ...picking whichever guides are closest to the transcription start site (TSS). We recommend targeting the...Church GM (2015) Highly efficient Cas9-mediated transcriptional programming. Nat Methods 12:326–328 . https...Nishimasu H, Nureki O, Zhang F (2014) Genome-scale transcriptional activation by an engineered CRISPR-Cas9 complex...

  14. Neuronal labeling with Spaghetti Monster

    Type
    Blog Post
    ...Spaghetti Monster probes at Addgene Uses of smFPs 1. Multi-channel connectomic tracers Fluorescent proteins... expanded use of smFPs in the future. References 1. Viswanathan, Sarada, et al. "High-performance probes...ideal probe should combine the solubility, cell tolerance, and optional fluorescence of FPs with facile ...

  15. What's Your Organism? Expanding Genomic Tools via the NSF EDGE Program

    Type
    Blog Post
    ...know that most cephalopods die after they lay their 1 clutch of eggs? Propagation is thus quite difficult...organism blog posts: Five Popular Model Organisms, Part 1 Five Popular Model Organisms, Part 2 ...species. Discovering ways to culture, propagate, transform, store, and share all of these new organisms continues...Phenotype thus making progress on the NSF “G to P grand challenge.” Creating a productive, new scientific...evolution because electric organs arose in different branches of the evolutionary tree at least 6-7 times and...mushrooms) but there are over 1,000 species. No transformation techniques are available yet. Polistes (Amy... a system using novel vesicles to efficiently transform these cells which don’t like foreign DNA at all...

  16. Plasmids 101: E. coli Strains for Protein Expression

    Type
    Blog Post
    ...folding, and deal with sub-optimal codon usage (Table 1) A few mutations are common to all or most expression...curated list of Bacterial Expression Systems Table 1: E. coli expression strains  Note: All strains are...specialized strains are available to confer greater transcriptional control, assist with proper protein folding...harboring this mutation are deficient in outer membrane protease VII, which reduces proteolysis of the...Lacks functional RNaseE which results in longer transcript half-life F- ompT lon hsdSB (rB- mB-) gal dcm...Chloramphenicol (pRARE) Good for “universal” translation; contains 7 additional tRNAs for rare codons ...pLemo (CamR) Expression of toxic, insoluble, or membrane proteins  T7 Express   IPTG-inducible expression...

  17. Site Directed Mutagenesis by PCR

    Type
    Blog Post
    ...sequence on either side of the desired mutation (usually 1-3 mismatched bases) is sufficient for your primers...researcher at Cornell University.       References 1. Breslauer, Kenneth J., et al. "Predicting DNA duplex...homology-directed repair (HDR) of a CRISPR/Cas9 induced double-stranded break. This site-directed genome editing requires...methylation-dependent endonuclease (i.e. DpnI), and bacteria are transformed with the nuclease-resistant nicked plasmid (the...products of the PCR reaction are pieces of double stranded, circular DNA with staggered nicks, recent data...final PCR products are pieces of linear, double stranded DNA with homologous ends. These homologous ends...the adenosine of the GATC recognition site. Transformation: After the PCR reaction, no ligation is required...

  18. Changing Labor Laws Bring Increased Postdoc Wages

    Type
    Blog Post
    ...$47,476 and the date for implementation was set for Dec 1 2016. In order to be exempted from the need to pay... these institutions (populations potentially from 1 to 5761, according to the NSF data) and therefore ...about? In March 2014, the White House issued a memorandum to the Secretary of Labor to update the FLSA,...postdocs in the final ruling was, for a lot of us, frankly quite surprising. Consortia of higher education...issued, a joint blog post by Director of the NIH Francis Collins and Secretary of Labor Thomas Perez, Fair...exactly how postdoctoral salaries are changing. Transparent salary information for postdoctoral positions... of postdocs at different points on this salary range. We have been able to gather data on changes to ...

  19. Prime Editing: Adding Precision and Flexibility to CRISPR Editing

    Type
    Blog Post
    ...possible without double-strand breaks (DSBs) or donor DNA templates.  Figure 1: An overview of prime...demonstrated ~20-50% efficiency in HEK293T cells with 1-10% indels. Since then, further innovations have continued...Engineering. https://doi.org/10.1038/s41551-024-01227-1 Yan, J., Oyler-Castrillo, P., Ravisankar, P., Ward... non-edited strand, they can cause the cell to remake that strand using the edited strand as the template...the nicked target strand. The initial result is a heteroduplex, with overlapping strands of edited and unedited...Improving reverse transcriptase efficiency Building upon prior reverse transcriptase research, the Liu...the edit into one strand, there’s a mismatch between the original sequence on one strand and the edited ...

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

    Type
    Blog Post
    ...different ways: Transformation grade DNA Cloning grade DNA Transfection grade DNA Figure 1: Transformation... differences between transformation, transduction, conjugation, and transfection Resources on Addgene.org...Transformation introduces transformation grade plasmids into a bacterial cell. Transformation grade DNA  Transformation...bacterial transformation of chemically and electrocompetent cells is minimal (picogram range). It can ...plasmids into mammalian cells? Use transfection grade DNA. Transfection is the process of inserting an ...without commercially available kits and DNA for transformation doesn’t need to be as pure as for other uses...scientists to clone from a smaller amount of DNA (ng range). To allow efficient and flawless cloning it is ...
Showing: 461 - 480 of 736 results