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We narrowed to 736 results for: RAN-1

Showing: 81 - 100 of 736 results

  1. TALEN Guide

    Type
    Collection
    ...coming closer to this DNA targeting ideal. Figure 1: (A) Representation of a zinc finger (ZF) nuclease..., that will probably never happen. Yet with transcription activator-like (TAL) effectors, scientists in...sequence-specific TAL effectors for modulating mammalian transcription. Nat Biotechnol. 2011 Feb;29(2):149-53. PMID...

  2. Getting the Most from Your Lentiviral Transduction

    Type
    Blog Post
    ...Check out Addgene's New Viral Service! References 1. Hay, R.J., Macy, M.L. & Chen, T.R. “Mycoplasma infection...contaminations.” Methods in Molecular Biology 946 (2013). 1-13. Pubmed PMID: 23179822. 5. Segura, M.M., Kamen,...Production, concentration and titration of pseudotyped HIV-1-based lentiviral vectors."Nature protocols 4.4 (2009...may not translate well to other cell types. To overcome this issue, try transducing with a range of dilutions...DEAE-dextran can improve transduction efficiency. In addition, this study found that transduction efficiency...passage number of a culture the lower the transfection and transduction efficiencies. Cultures should only be...others opt for a single harvest after transfecting with the transfer and packaging plasmids. A benefit of...

  3. Save Time with Transient Plant Leaf Transformations

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    Blog Post
    ...regulated in the medicinal plant, the Catharanthus roseus (Figure 1). The ultimate goal of our research ...reports 9.1 (1990): 1-5. PubMed PMID: 24226366. Lee, Min Woo, and Yinong Yang. "Transient expression assay... plant scientists do transient leaf transformations The development of transgenic plants is a common problem...that the transformation and gene expression can be tracked. During a transient leaf transformation, the engineered...method. Transfer your plant into the dark 16 hours before the transformation. We routinely transfer our plants...developing transgenic hairy root lines in tissue cultures takes half a year, and generating a transgenic plant...plant can take even longer, a transient plant leaf transformation process could save the plant biologist...

  4. CRISPR Plasmids - Plants

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    ...Hyg Chen 86196 pYPQ141-ZmUbi-RZ-As Maize ubiquitin 1 none As Cpf1 Qi 91715 pKEE401 yes, cut S. pyogenes...introduce gRNA-targeted single-strand breaks in DNA instead of the double-strand breaks created by wild type... opposite strands of your DNA in close proximity. These double nicks create a double-strand break (DSB...dead dCas9 fused to a transcriptional activator peptide can increase transcription of a specific gene. ...Genome Editing Cut Base Edit Nick Prime Edit Transcriptional Regulation Activate Interfere Epigenetics RNA...functional CRISPR/Cas enzymes will introduce a double-strand break (DSB) at a specific location based on a gRNA-defined...base editors convert C->T (or G->A on the opposite strand) within a small editing window specified by the...

  5. CRISPR 101: Any Base Transversion Editors

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    Blog Post
    ...glossary at the end of this post! Figure 1: Base transitions are edits between the purine bases (adenine...uracil DNA glycosylase. Nature Communications, 15(1), 4897. https://doi.org/10.1038/s41467-024-49343-5...uracil-DNA glycosylase. Nature Communications, 15(1), 6397. https://doi.org/10.1038/s41467-024-50012-w... the non-edited strand to encourage the cell to repair the DNA using the edited strand as a template —...editing. Like other transversion editors, this editor had relatively high indel rates ranging from 4.7% to 30%...our last post, we talked about the first base transversion editors: CGBEs, or C → G Base Editors. CGBEs...editor could excise other bases? Could other transversion editors be close at hand? Once again, this post...

  6. New Optogenetic Tools for Cytoskeleton and Membrane Control

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    Blog Post
    ...overcomes many of those limitations (Figure 1). Figure 1: Opto-katanin induces fast, localized microtubule...Signaling Toolbox. ACS Synthetic Biology, 11(1), 515–521. https://doi.org/10.1021/acssynbio.1c00604.  Mahlandt...specific area at the plasma membrane (Figure 2). Just express the plasma membrane-localized iLID along with...cell membrane isn’t powerful enough, what about altering the chemical composition of the membrane itself...SuperPLD hydrolyzes membrane PC to PA. A) OptoPLD is recruited to plasma membrane or lysosomes and its...optogenetic tools to control the cytoskeleton and cell membrane dynamics. First, a quick refresher.  The tools...undergoes a conformational change and binds the plasma membrane under blue light CRY2-CIB1 heterodimerizes or ...

  7. Viral Production

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    ...to determine purity of the AAV preparation. Figure 1: Silver staining of purified and non-purified AAV ...of Pennsylvania Vector Core. Transfections are performed using the transfer plasmid, a plasmid encoding...cell cultures are inspected for sterility. Transducibility Some viral vectors are tested in vitro and ...section for images). Figure 2: AAV Pro cells were transduced with either pAAV-Ef1a-mCherry-IRES-Cre (55634...filtration through a 0.45 µm polyethersulfone (PES) membrane. The preparations are then aliquoted, frozen, ...opens in a new window) ). Briefly, 293T cells are transduced with serial dilutions of a lentiviral vector,... taken off-line and discarded. Confirmation of Transfer Plasmid Addgene uses a rigorous barcode matching...

  8. R Bodies: Membrane-Rupturing Microscopic Tools

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    Blog Post
    ...whether they could 1) be decorated with proteins and remain functional, 2) break membranes outside of their...Follow her on Twitter @jessicapolka.     References 1. Kanabrocki, J. A., Quackenbush, R. L. & Pond, F. ...learn a lot from Nature’s simpler solutions to transmitting forces over long distances. For instance, how..., R bodies do not exert force by dynamically rearranging into different polymerization states. Instead...demonstrates that R bodies can function to break membranes of different organisms. And finally, we demonstrated...could use them to lyse cells (and surrounding membranes) in keeping with their natural function (see below...

  9. Targeting HIV-1 with CRISPR: Shock and Kill or Cut it Out?

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    Blog Post
    ...plasma HIV-1, improving life expectancy and quality for HIV-1 patients. However, ART is not an HIV-1 cure. ...activated HIV-1, measured via HIV-1 promoter-driven GFP expression. In cell lines that can produce HIV-1 toxic...with HIV-1. For two HIV-1 strains, Cas9/gRNA lentiviral expression significantly reduced the HIV-1 copy number...HIV-1 transcription and destroy infected cells, the other using wild-type Cas9 to remove the HIV-1 genome...currently infected with the lentivirus HIV-1. Today, HIV-1 can be controlled with antiviral therapies ...take on HIV-1 in vitro, and what obstacles must be overcome for clinical success. ART and HIV-1 reservoirs...Cutting HIV-1 out of the genome with CRISPR/Cas9 Before the introduction of CRISPR, HIV-1 genome editing...

  10. Transferable Skills Guide: Cross-team Communication

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    Blog Post
    ...Addgene, we use seven different types of “plates” (Fig. 1). When you have to tell an engineer that you want ...”? We took steps towards solving this problem by 1) making sure we qualified these words when we discussed...resources on the Addgene blog Read about other transferable skills Find all of our science career related...

  11. 5 Steps to Writing Grant Proposals for Grad Students

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    Blog Post
    ...offers insight into a career as a Professor. Step 1: Your PI forwards you a Request For Applications (...proposal typically lists the deliverables, explains 1-2 potential pitfalls of the intended protocol, and...improve your grant writing skills. Good luck! Further advice on grant writing Since successful grant writing... during the grant application process. When my PhD advisor first asked me to write a grant application... advice on writing a grant proposal. "Murder Most Foul: How Not to Kill a Grant Application" by Vid Mohan-Ram...their advisor alone applies for these competitive grant applications; however, in some cases, your advisor...advisor may choose to allot some if not all grant duties to trusted students. For example, if the proposal...

  12. Typing CRISPR Systems

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    Blog Post
    ...   Class 1 versus class 2 There are two CRISPR classes: Class 1 and Class 2. Class 1 CRISPR systems...archaea, though Class 1 remains largely underutilized compared to Class 2. Class 1 Despite their incredible...optimal plasmids to harness the power of Class 1 systems. Class 1 CRISPR systems can be further broken down...6105. https://doi.org/10.1093/nar/gku241 Cong, L., Ran, F. A., Cox, D., Lin, S., Barretto, R., Habib, N....types, 33 subtypes, and 17 variants. Figure 1: Overview of the CRISPR classification hierarchy. ...effector complex after binding to a target. Class 1 is the most abundant of the two, comprising about ...incredible abundance, Class 1 CRISPR systems are less popular in the lab. This can be attributed to the multi-Cas...

  13. 6 Tips for Grant Writing

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    Blog Post
    ...improve your grant writing and hopefully increase your success rate. 1. Submit your grant early! First...involved so it’s important not to throw in the towel if 1 or 2 funding bodies say no. If possible, get feedback...in writing small travel grants to attend conferences and a seed funding grant for a proof-of-principal...www.assaygenie.com. No matter what facet of academia you are in, grant writing can be the lynch pin that results in your...to be much use. 2. Originality is essential for grant success The two most important things reviewers ...sure that comes across. Check out some in-depth grant writing tips at the Human Frontier Science Program's...Skills 5. Practice makes perfect, apply for small grants too There are actually quite a lot of different...

  14. No Llamas Required - Synthetic Nanobodies Against Membrane Proteins

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    ...crevices proteins on the cell surface. Figure 1: Comparison of common antibody, HCab, and nanobody...Humana Press, pp 133–148. https://doi.org/10.1007/978-1-62703-293-3_10 Walter J et al. (2020) Synthetic nanobodies...sybodies against membrane proteins Developing antibodies against proteins embedded in membranes is tricky because...develop antibodies against unstable targets like membrane proteins that may unfold due to camelid high body...synthetic nanobodies, called sybodies, against membrane protein targets in as little as 3 weeks (Zimmermann...lab can quickly select for sybodies targeting a membrane protein of interest using the Sybody Generation...have a concave, loop, or convex scaffold and are randomized with a defined set of different trinucleotides...

  15. When Fidelity Matters: A frank discussion about ligase fidelity

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    Blog Post
    ...phosphorylated DNA termini to 3 ́-OH DNA termini (1-4). In vitro, ligases (notably T4 DNA ligase) are ... enzymology of DNA repair enzymes.     References 1. Shuman, Stewart, and Christopher D. Lima. "The polynucleotide...thermostable DNA ligase-encoding gene." Gene 109.1 (1991): 1-11. PubMed PMID: 1756968. 7. Lauer, G., et al. "Cloning...19329793. PubMed Central PMCID: PMC2719376. 5. Barany, Francis. "Genetic disease detection and DNA amplification...1986365. PubMed Central PMCID: PMC50775. 6. Barany, Francis, and David H. Gelfand. "Cloning, overexpression...Central PMCID: PMC208194. 8. Luo, Jianying, and Francis Barany. "Identification of essential residues in Thermus...(1989): 245-254. PubMed PMID: 2753355. 12. Barany, Francis. "The ligase chain reaction in a PCR world....

  16. pCXLE toolkit: Efficient episomal plasmid-based method to reprogram peripheral blood cells to iPSCs

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    Blog Post
    ...expressing additional EBNA-1 transiently. The transiently expressed EBNA-1 increases protein expression...are based on the Epstein-Barr Nuclear Antigen-1 (oriP-EBNA-1) system where each vector contains a viral ...replication, OriP and the EBNA-1 sequence that encodes for the DNA binding protein EBNA-1 [7]. These two sequences...extra plasmid, pCXWB-EBNA-1 is used in the reprogramming cocktail. pCXWB-EBNA-1 lacks oriP and is unable...pCXLE vectors listed below in Table 1 are available at Addgene. Table 1: List of pCXLE episomal plasmids,... and shRNA against p53 pCXWB-EBNA1 37624 transient EBNA-1   Many thanks to our guest blogger, Kusumika...obtained and the subsequent reprogramming success [1]. Fibroblasts need to be cultured and expanded in ...

  17. Split Fluorescent Proteins for Studying Protein-Protein Interactions

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    Blog Post
    ...between strand 10 and 11 to yield large (1–10) and small (11) fragments. In several cases an FP(1–10) fragment...General Design Figure 1: Two proteins are fused to the FP(11) and FP(1–10) fragments. When Protein... to Plasmids) PI Blue EBFP2 EBFP2(1–10) and Capri(1–10) for use with GFP(11) Multiplexed labeling...Commun. 2017 Bo Huang mNeonGreen3 mNG3K(1–10) and mNG3A(1–10) for use with mNG2(11) Improved yellow-...sfCherry2 sfCherry2(1–10) and sfCherry2(11), also photo-activatable variant PAsfCherry2(1–10) Improved split...brings the pieces of the FP back together (Figure 1). This method is generally known as Bimolecular Fluorescence...even from a different base FP (for example, EBFP2(1–10) can be used with GFP(11)). Some of the articles...

  18. Neurodegeneration Research Collection

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    ...one of three different inherited genes: Presenilin 1, Presenilin 2, and APP gene.The majority (>90%) of...used to transform iPSCs and fibroblasts into neurons and more. New and Noteworthy: Study aberrant axon initial...associated with the disease and implicate a wide range of cellular processes. The vast majority of cases...kinetics and localization for imaging synaptic transmission. Aggarwal et al. Nat Methods. 2023 May 4. See...PiggyBac plasmids with tet-inducible expression of transcription factors for sensory, motor, and cortical neuron...Use a FRET-based method to monitor structural transitions in protein self-organization. Wan et al. Cell...

  19. CRISPR/Cas9 FAQs Answered!

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    Blog Post
    ...probably have better specificity. A good reference is Ran et al. (2013). As for target selection, we usually...fine based on new data). For general transfection, you could use 1 µl of 10 µM oligo stock for each well...well in 24-well plate. A good example/reference is Ran et al. (2013). In the case where you have intact...https://doi.org/10.1534/g3.112.005439  Cong, L., Ran, F. A., Cox, D., Lin, S., Barretto, R., Habib, N.... from Double-Strand Break Repair in Mammalian Cells. Molecular and Cellular Biology, 18(1), 93–101. https...https://doi.org/10.1128/mcb.18.1.93  Ran, F. A., Hsu, P. D., Lin, C., Gootenberg, J. S., Konermann, ...1389. https://doi.org/10.1016/j.cell.2013.08.021  Ran, F. A., Hsu, P. D., Wright, J., Agarwala, V., Scott...

  20. Quick, Versatile Plant Transgenesis with GreenGate Plasmids

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    Blog Post
    ...making it easier for us to do our work.” References 1. Athanasios Lampropoulos et al. "GreenGate - A Novel...terminators and resistance cassettes for selection of transgenic plants. The complete GreenGate toolkit, now available...the time required to generate multi-construct transgenic plants. In the most practical terms, what GreenGate...Versatile and Efficient Cloning System for Plant Transgenesis." PLoS ONE. 8(12): e83043 (20 December 2013)...
Showing: 81 - 100 of 736 results