We narrowed to 659 results for: Clu
-
TypeBlog PostPublishedOct. 13, 2022, 1:45 p.m....domain - VP64. Later generations of activators include the SunTag system which facilitates recruitment...Addgene’s website. Fig. 3: Cas9 repressors include dCas9 alone and a dCas9-KRAB fusion, both targeted...targeted to the promoter region. Cas9 activators include Cas9 fused to VP64, VP64 along with other SAM factors...expression in different species and cell types including (but not limited to) bacteria, yeast, plants, ...
-
New CRISPR Tools: Cas7-11 and PASTE
TypeBlog PostPublishedFeb. 17, 2022, 2 p.m.... hardest part of any revolutionary discovery, including CRISPR, is turning potential into impact. In molecular..., PhD, and Jonathan Gootenberg, PhD, focuses exclusively on this space — developing tools for cellular...measurable cell toxicity – using Cas7-11. This included tests done in cell lines known to be more sensitive... — the ability to now make any small changes, including base edits, insertions, and deletions. But CRISPR... -
Antibodies 101: Reading a Flow Plot
TypeBlog PostPublishedFeb. 6, 2024, 3:13 p.m....an X,Y axis to generate a dot plot (Figure 1). Clustered events are called a population. ..., like a box, ellipse, or circle, drawn around clusters to define populations. This shape is called a ...diameter and volume of the cell, thus allowing the exclusion of doublets and debris. The side scatter roughly...show frequency of events, instead of relying on clustered dots. In a contour plot, each line contains the... -
New Acoustic Reporter Genes: Ultrasound Imaging of Gene Expression
TypeBlog PostPublishedJan. 24, 2023, 2:15 p.m....inside the body. So we screened gas vesicles gene clusters from a diverse set of gas vesicle-expressing species...right: colony whiteness on inducer plates, white inclusions in cells with GVs via phase contrast microscopy...this was the Anabaena flos-aquae gas vesicle gene cluster because of its small size and its ability to produce...a time-intensive endeavor. That is why we also included an analogous set of transient constitutively-expressed... -
Important Considerations in Optogenetics Behavioral Experiments
TypeBlog PostPublishedOct. 1, 2015, 2:30 p.m....but some examples I’m personally interested in include: intracranial self-stimulation (ICSS) and place...resolve these issues is with electrophysiology. Concluding thoughts and moving beyond the mouse While it...applicable to the rat [6]. Technical differences include the ferrule implants and patch cables. One huge... at the Rockefeller University. His interests include the role of learning and memory in opioid addiction... -
Fluorescent Proteins 101: GFP Fusion Proteins - Making the Right Connection
TypeBlog PostPublishedApril 9, 2019, 1:13 p.m....known as steric hindrance. For instance, GFP may occlude catalytic sites or obstruct binding sites or motifs...proteins that behave as monomers in mammalian cells include: mTurquoise2, mEGFP, mNeonGreen, mScarlet(-I), ...motif, which is needed for Golgi localization, is occluded and therefore this protein is mislocalized. The...that the fusion behaves well. But this is not conclusive evidence that the fusion operates in the same... -
Fluorescent Proteins 101: When GFP lets you down
TypeBlog PostPublishedAug. 23, 2018, 12:05 p.m....Several factors contribute to the popularity of GFP including (i) fast and complete maturation to functional...endogenous fluorescent co-factors can be used. Examples include the LOV domains that bind flavin (Buckley et al... conditions, with pKa values below 4. Examples include mTurquoise2 (pKa = 3.1), tagRFP (pKa = 3.1) and... or media. These sources of ‘autofluorescence’ include riboflavin in the medium or flavin containing proteins... -
Evolution of Brainbow: Using Cre-lox for Multicolor Labeling of Neurons
TypeBlog PostPublishedApril 24, 2015, 2:39 p.m....works. Advances in Cre-lox recombination have included inducible Cre, such as the tamoxifen-responsive...mediate recombination.) The deletions are mutually exclusive, as either deletion removes one of the other loxP...immediately following the promoter. The STOP cassette includes a mutant YFP that does not fluoresce, but can ...More variants have been created by other labs, including R26R-Confetti described in Hugo J. Snippert, et... -
Finding Your Science Policy Path
TypeBlog PostPublishedSept. 22, 2020, 1:15 p.m....organizations in a variety of science policy roles including fellowships, volunteer positions, and part-time... with a large set of other societal pressures including economic, cultural, and political. Science policy...science blogs. Other avenues for skill development include leadership of extracurricular organizations, working...opportunities. NSPN maintains a great resource guide including conferences, societies, and outreach groups. Grab... -
Antibodies 101: Validation
TypeBlog PostPublishedMarch 24, 2022, 1:15 p.m.... molecule at all, leading to inconclusive results, erroneous conclusions, and wasted time. In fact, many...antibody-dependent approach. Antibody-independent methods may include analysis of transcriptomic or proteomic data from...specifically to the expected protein. Common approaches include using RNAi to knock-down expression of your target... -
Sequencing Options for CRISPR Genotyping
TypeBlog PostPublishedOct. 4, 2016, 2:30 p.m....for CRISPR genotyping. The most common options include mismatch cleavage assays, such as Surveyor™, T7E1...information profile regarding putative off-target sites including both the edit rate and the repair product sequence...algorithms do not take into account other elements, including those relating to DNA structure (e.g. epigenetic...options exist for unbiased off-target detection, including Digenome-seq (Kim et al. 2015) for in vitro analysis... -
Fluorescent Proteins 101: Monitoring Cell Mobility Using Fluorescent Proteins
TypeBlog PostPublishedAug. 15, 2017, 1:24 p.m....organism to make the complicated transition from a clump of cells to a fully differentiated body. In contrast...monitor cell interactions in diverse tissues, including lymph nodes, brain, liver, gut and skin. Two-...able to help answer using two-photon microscopy include: How do T cells and/or Neutrophils travel to the...probes when using a standard Ti:Sapphire Laser include: Blue-green fluorescent proteins like EGFP Yellow-orange... -
Antibodies 101: Introduction to Antibodies
TypeBlog PostPublishedJan. 19, 2021, 2:15 p.m.... Figure 1: Labeled diagram of an antibody including Fc, Fab, heavy chain, light chain, constant region...antibodies for experiments in a variety of fields including neuroscience and immunology. They can be used ...analyzed using techniques such as flow cytometry. Conclusion Whew! Now you know that antibodies can be used...used for. Addgene’s Antibody Plasmid Collection includes plasmids that express antibodies, nanobodies, ... -
Typing CRISPR Systems
TypeBlog PostPublishedMarch 18, 2025, 1:15 p.m.... effectors and are the only CRISPR type that exclusively targets RNA. Cas13 has become a natural successor...CRISPR will likely depend on a variety of factors, including your goal, experience, hypothesis, and even the...keep increasing, as efforts using large-scale clustering and discovery algorithms find new candidates ...of rare CRISPR-Cas systems with deep terascale clustering. Science, 382(6673). https://doi.org/10.1126/... -
A Primer on Optogenetics: Introduction and Opsin Delivery
TypeBlog PostPublishedSept. 10, 2015, 2:30 p.m....main experimental avenues (that are not mutually exclusive) commonly used in optogenetics: 1) electrophysiology.... Of course, proper controls are required and include, for example, confirmation of opsin expression ... brief, some standard opsins (see table above) include Channelrhodopsin2 (Chr2), a blue-light sensitive... at the Rockefeller University. His interests include the role of learning and memory in opioid addiction... -
Participate in the Pets of Addgene Scavenger Hunt
TypeBlog PostPublishedApril 1, 2021, 1:15 p.m.... this scavenger hunt and the subsequent clue. (Begin with clue 1 below!) You will find five photos this... the blog. Below each photo, the pet gives you a clue to help you find the next picture. If you find the... -
CRISPR 101: Multiplex Expression of gRNAs
TypeBlog PostPublishedSept. 10, 2020, 11:45 a.m....for plasmids suitable for other model systems, including E. coli, plants, Drosophila, and zebrafish! ...destination vectors. Destination vector options include humanized wt Cas9, dCas9 (transcriptional repressor... Figure 3: Comparison of Multiplex Strategies including Standard PolIII-gRNA cassette, Csy4-cleavable ...filler sequences. You also have the option of including a previously-designed tyr gRNA, which causes hypopigmentation... -
Selecting Your Plasmid Purification Kit
TypeBlog PostPublishedMay 27, 2025, 1:15 p.m....sneak in from almost any step of the process and include things like genomic DNA, RNA, proteins, salts, ...sustainable practices for the lab. Plastic consumables (including pipette tips, plates, and yes those purification... kits from a variety of manufacturers. It also includes a rough comparison of list price between these...Yeast, BAC, transfection-grade While we’ve included a sample of some commonly used kits, please note... -
CRISPR Cheat Sheet
TypeBlog PostPublishedJan. 27, 2021, 2 p.m....new edition of the eBook includes the details on advances in CRISPR including: Base editing Prime editing...CRISPR Cheat Sheet here! CRISPR Cheat Sheet Clustered Regularly Interspaced Short Palindromic Repeats... -
Plasmids 101: How to Name Your Plasmid in 3 Easy Steps
TypeBlog PostPublishedFeb. 13, 2014, 1 p.m....for plasmid. pXXXXX-XXXX Step 1: Backbone name Include the empty backbone name in your plasmid name. This...backbones, visit Vector Database. Step 2: Insert name Include information about the insert in your plasmid name...contains a mutation or modification, this should be included in the plasmid name. Mutations are generally listed...