We narrowed to 208 results for: PHI-1
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TypeGuide... most popular genome engineering approach. Figure 1: Overview of the basic CRISPR mechanism Engineered...from Staphylococcus aureus ) has a coding sequence ~1 kb shorter than SpCas9 while retaining the same basic...lentiviral transfer vectors (Figure 8B). Libraries come in 1-vector systems, in which Cas9 is included in the gRNA-containing...efficiency and fidelity. Nature Communications , 13 (1), 1425. PMID: 35301321 Edraki, A., Mir, A., Ibraheim...interrogation by SpRY-Cas9. Nature Communications , 15 (1), 3663. PMID: 38688943 Hsu, P. D., Scott, D. A., Weinstein...increase its specificity. Nature Communications , 9 (1). PMID: 30082838 Maddalo, D., Manchado, E., Concepcion...CRISPR-Cas9 with Bacteriophage Proteins. Cell , 168 (1–2), 150-158.e10. PMID: 28041849 Sakuma, T., Nishikawa...
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Bacterial Expression Systems
TypeCollection...expression inducible by red light. pCph8 50552 pLtetO-1 Anhydrotetracycline Jeffrey Tabor Anhydrotetracycline... pZE31ms2 25860 MS2 coat protein binding sites Philippe Cluzel RNA localization: Contains a 2x array of...DsRed-ms2x2 26248 MS2 coat protein binding sites Philippe Cluzel RNA localization: Contains a 2x array of...Fluorescent protein, GUS, and LacZ Derivatives avaliable Philip Poole A family of promoter probe vectors incorporating... -
Viral Vectors 101: Systemic Capsids
TypeBlog Post...total dose between 1 x 1011 and 5 x 1011 vg per animal (i.e., 10 and 50 µL of vector at 1 x 1013 vg/mL titer... and 1 x 1012 vg*. Most systemic capsids are easy to produce in high titer, but higher doses (>1 x 1012...before using it in an application. Figure 1: AAV toolkit for targeted gene delivery to the brain...decision tree for selecting a capsid, while Table 1 gives a brief overview of published descriptions of...Capsids page to learn more about that collection. Table 1: Systemic capsid plasmids available from Addgene ...AAV9. At lower viral titers than AAV-PHP.B (e.g., 1 x 1011 vg/mouse), AAV-PHP.eB can transduce a greater...Diverse Brain Cells. Annual Review of Neuroscience, 45(1), 447–469. https://doi.org/10.1146/annurev-neuro-111020... -
Crowdfight, a Platform to Boost Scientific Collaboration During COVID-19 and Beyond
TypeBlog Post...prompted us to create Crowdfight. Figure 1: Crowdfight is a community of scientists who believe...collaborator for a specific task, Crowdfight will: 1) Find an expert with the right skill; 2) Confirm the...volunteers helped with things like translations, bibliographic searches, etc. Expanding collaborations... -
10 Steps to a Perfect Science Talk
TypeBlog Post...Resist the urge to say everything I allow 1 slide for every 1 minute of talk length, and then I cut a few...NatureJobs: Interviews: The All-Important Job Talk. 1. Watch other people give great talks We are not talking...Like graphing data, choosing controls, or mixing clear solutions—public speaking is skill that any scientist... -
New FLIS Plasmid (April Fools 2022)
TypeBlog Post...for locating and containing escaped flies. Nature 4:1-22. https://doi.org/4.01.2022/hafd...FLIS and Flies Drosophila, more commonly known as fruit flies, have been an important animal model for... -
Plasmids 101: Inducible Promoters
TypeBlog Post...Hsp70, the heat shock releases heat shock factor 1 (HSF-1), which subsequently binds to heat shock elements...genome engineering in species like C. elegans and Drosophila. Light inducible promoters Light is another way... -
Our Most Popular Blog Posts from 2019
TypeBlog Post...deletions, and all possible base-to-base changes. 1. Plasmids 101: Transformation, Transduction, Bacterial...Scientific Graphics by Beth Kenkel Find a round-up of tools you can use for making scientific graphics. PS:... -
CRISPR 101: Multiplex Expression of gRNAs
TypeBlog Post...post about multiplexing with Cpf1. Figure 1: Multiplexing allows researchers to express multiple...be used in a wide range of bacteria. References 1. Maddalo, Danilo, et al. “In vivo engineering of oncogenic...other model systems, including E. coli, plants, Drosophila, and zebrafish! Why use multiplexed gRNAs?...restriction enzyme BsaI. If you’re working in Drosophila, a two-gRNA expressing plasmid is available from...supplied on a separate plasmid. Multiplexing in Drosophila Bullock Lab Multiplex Plasmid: Two gRNAs can ...or SLIC cloning. gRNAs are expressed from two Drosophila U6 promoters. Cas9 must be supplied on a separate...efficient germline and somatic genome engineering in Drosophila.” Proceedings of the National Academy of Sciences... -
28 Hot Plasmid Technologies from 2015
TypeBlog Post...light-sensitive LOV2 domain of Avena Sativa phototropin 1 (AsLOV2). In the dark, this peptide is caged by the...iLID nano and 800nM to 47µM for iLID micro; Figure 1). With this great affinity range, these LIDs have ...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... al., BMC Molecular Biology. 2013 August 20th; 14(1). TRICK: A method for visualizing the first round...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... -
Viral Vectors 101: Biosensors
TypeBlog Post...Introduction to Biosensors. Essays Biochem. 2016 Jun 30;60(1):1-8. DOI: 10.1042/EBC20150001 Leopold, A. V., Shcherbakova...of the biosensors used in research are very sophisticated, but at their core they can all be boiled down... -
Back to Bacteria: CRISPR gRNA Multiplexing Using tRNAs
TypeBlog Post...modified slightly with either a 1-7 base poly(U)tail in tRNA-gRNA constructs or a 1-4 base tail from the second...three strategies described in this post in the graphic below - including some simple comparisons. The... -
Antibodies 101: Introduction to Gating in Flow Cytometry
TypeBlog Post...at a low angle (FSC) or large angle (SSC) (Figure 1). FSC values depend on the cell's size, while SSC ... or ask experienced colleagues. Figure 1: The laser pulse illuminates the cell and is scattered...and W describe the shape of a histogram, which graphically represents the time and intensity of the cell...wondering which second measurement to choose for graphically displaying your events. You are free to choose... -
Gendered Innovations: Why Does Sex of the Cell Matter?
TypeBlog Post...healthcare system was $35.2 billion (or $140 per $1 invested in the trial). The goal of gendered innovations...is the goal of Gendered Innovations. This project: 1) develops state-of-the-art methods of sex and gender... work to be done! Researchers need to learn sophisticated methods of sex and gender analysis. Universities... -
Evolution of Lab Techniques
TypeBlog Post...blotting, named for its inventor Edward M. Southern (1), can be used to detect DNA identity, size, and abundance...hours at low cost with restriction mapping (Figure 1). Though some labs may prefer to send DNA directly... serotonergic system and behavior. References 1. Southern, Edwin Mellor. "Detection of specific sequences...simian virus 40 DNA by restriction endonuclease of Hemophilus influenzae." Proceedings of the National Academy... -
Molecular Biology Reference
TypeGuide...Invitrogen F- Phi80lacZDeltaM15 Delta(lacZYA-argF) U169 recA1 endA1 hsdR17(rk-, mk+) phoA supE44 thi-1 gyrA96...) xyl-5 λ– leu mtl-1 Top10 Invitrogen F- mcrA Delta(mrr-hsdRMS-mcrBC) Phi80lacZM15 Delta-lacX74 recA1 ...Addgene; Promega e14-(McrA-) recA1 endA1 gyrA96 thi-1 hsdR17(rK- mK+) supE44 relA1 Delta(lac- proAB) [F ...use, dilute your antibiotic into your LB medium at 1:1,000. For example, to make 100 mL of LB/ampicillin...Invitrogen F- mcrA Delta(mrr-hsdRMS-mcrBC) Phi80lacZDeltaM15 Delta-lacX74 recA1 araDelta139 D(ara-leu)7697... -
How Addgene Creates Content
TypeBlog Post...to do that than in the Addgene blog? Fig. 1: One of our favorite content collections is our Help...of text. Since our guides are heavily used, our graphic designer made most of the Antibody Guide figures...communication between our content team and our graphic designer to make sure things are clear, accurate...Fig. 2: Reference image (left) provided to our graphic designer upon request that the science communicator...design (right). Immediate and immense regret of the graphic designer not shown. However, for our blog... -
CRISPRainbow and Genome Visualization
TypeBlog Post...all the colors of the CRISPRainbow. Figure 1: CRISPRainbow was used to track the localization of...is available as a kit from Addgene. References 1. Chen, Baohui, et al. “Dynamic Imaging of Genomic ...this rainbow of colors, Ma et al. performed sophisticated tracking of multiple chromosomal loci in live... -
CRISPR 101: Cas9 vs. The Other Cas(s)
TypeBlog Post...The main differences between Cas7-11 and Cas13 are 1). non-specific RNA cleavage and 2). cell toxicity.... targeting single stranded RNA. Table 1: Fast facts on Cas family proteins and genome editing...to detect variants such as single nucleotide polymorphisms. Fast facts – PAM requirement: none. Best ... -
Hot Plasmids and Viral Preps - March 2021
TypeBlog Post...in a “single pot.’ Figure 1: Single-Pot Assembly: a) Graphic depiction of chromatin within a nucleus...AID system relies on a transport inhibitor response 1 (TIR1) expressed under a desired promoter with a specific...