We narrowed to 735 results for: RAN-1

...one of three different inherited genes: Presenilin 1, Presenilin 2, and APP. The majority (> 90%) of individuals...associated with the disease and implicate a wide range of cellular processes. The vast majority of cases...deposition with APOE2 constructs . (Hudry et al. Sci Transl Med. 2013) Express human DNMT1 with a Myc tag. ...vesicular brain cells and more. Measure synaptic transmission with improved glutamate indicators . (Aggarwal...Differentiation Factors Find plasmids used to transform iPSCs and fibroblasts into neurons and more. Convert...plasmids expressing SOX10, OLIG2, and NKX6.2 transcription factors for rapid and efficient generation of...astrocytes to dopaminergic neurons using three transcription factors in a single polycistronic lentiviral...

...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...

...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...

...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 ...

...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...

...infection to host cells . Signal transduction and targeted therapy, 5 (1), 283. https://doi.org/10.1038/...envelope protein, E; nucleocapsid protein, N; and membrane protein, M), including plasmids for protein production...Anderson et al., 2020). BSG plasmids - (CD147), transmembrane glycoprotein of the immunoglobulin superfamily...proteolysis of SARS-CoV Spike protein, activating its membrane fusion function (Bosch et al., 2008). TACE plasmids... protease (Mpro) activity reporters Reverse transcriptase, protease, and polymerase constructs for SARS-CoV...Llamas Required - Synthetic Nanobodies Against Membrane Proteins Return to top Pooled Libraries Pooled...

...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...

...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...

...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...

...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...

...   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...

...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...

...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....

..., 2.4 MB Zhang GeCKO pooled library amplification 1 vector system: lentiCRISPR v2 2 vector system: lentiCas9...sequence and checks for off-target binding. Supports a range of Cas enzymes (Cas9 and Cas12) and organisms. Developed...gene inactivation, NHEJ, and HDR. Includes a wide range of reference genomes, including animals, plants,...and cytosine (CBE) base editors. Includes a wide range of reference genomes, including animals, plants,...Ui-Tei Lab . CRISPick (Link opens in a new window) Ranks and picks candidate CRISPRko/a/i sgRNA sequences...for off-target binding. Currently supports a wide range of organims. Developed by the Michael Boutros Lab...and Wente Zebrafish: gRNA cloning, in vitro transcription, injection gRNA core ; Cas9 ; optimized Cas9...

...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 ...

...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...

...and placed in a buffer. Antibody Structure Figure 1: Structure of an antibody A standard antibody is made... together to form a “Y” shape, as shown in Figure 1. The two arms of the Y structure are responsible for...proteins by size. Transfer proteins from the gel to a membrane. Incubate the membrane with an unconjugated...block the membrane to remove excess antibody and prevent unwanted binding. Incubate the membrane with a ...Wash and block the membrane. Activate the HRP with a substrate kit. Image the membrane using either X-ray...hybridoma cells into plasmids. Plasmids are then transfected into cells and induced to express antibodies....variable regions, shown in light blue and light orange, that bind to a specific antigen, triggering an...

...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...

...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)...

...just make their own buffers when the kits buffers ran out. Homemade buffers performed the same as those...series to increase binding capacity. Figure 1: Purify DNA using glass filters. Image: Kim and Morrison...a gravity-based column plasmid maxiprep kit takes 1-1.5 hours to complete, while this syringe-based method... solution. NaI has two purposes in this protocol: 1) it solubilizes both DNA and agarose, and 2) it helps...property and developed a protocol that uses Whatman No. 1 paper or even paper towels to purify DNA and RNA from...rather a glass powder slurry that has a milk-like appearance. To make DNA stick to the glass particles in ...the Patterson Protocol was shown to purify DNA ranging in size from 100 bp to 48 kb with high yields and...