...the green “Save Verified Data and Return to Article” button to return to the table listing all of the plasmids...the full plasmid sequence can be assembled or theoretical — it does not have to be entirely verified by...recombination in bacteria) such as lentiviral, retroviral, and AAV plasmids, we recommend the NEB Stable...process for all of the plasmids in your deposit, return to the Add Plasmids page and click the green “I...

...Combinatorial strategies can also be used for retrograde tracing: visualizing neurons that received information...region. In the example shown in Figure 1, injected retrograde AAVs at one location result in opsin expression...viral vector related blog posts Learn more about retrograde AAVs Resources on Addgene.org Read our AAV ...guide Learn more about our viral service Find retrograde AAVs at Addgene ...

...principle of bioluminescence resonance energy transfer (BRET). Photons emitted by a Renilla luciferase variant...This technique is very similar to the widely used FRET, but it eliminates the need for an excitation light...exhibited 3-5 fold increased brightness over previous BRET probes, with sufficient signal for in vivo and in...Check Out All Our Fluorescent Protein Resources Find FRET Plasmids Find Empty Backbones with Fluorescent Protein...

... be fused to other proteins, and these fusions retained the ability to bind GFP. The idea of using GFP...T-DDOGs, GFP, and a reporter construct into the mouse retina. GFP expression robustly activated the reporter...Cre-DOG is both robust and specific. When tested in retinal electroporation studies, DsRed reporter expression...studies, and since cells infected with AAV-Cre-DOG retain normal neuronal function, this system may make ...

...cause an effect called FRET (fluorescence [or Förster] resonance energy transfer). FRET occurs when energy...excites the fluorescence of another FP (e.g. YFP). FRET only occurs when the distance between the two FPs...considered when labeling proteins that interact. Indeed, FRET is often used to determine if two proteins interact...

...and stop. For instance (chr3, 1, 10) indicates a stretch of DNA at the very beginning of the third chromosome... Let’s now examine how we can locate powerful stretches of DNA known as enhancers with the help of genome...ESCs) with a lentiviral vector, an attenuated retrovirus in the same family as HIV. By design, our lentivirii...light on dark matter Precise knowledge of which stretches of the genome are pillars for stem cells, or metastasized...

...domain being tested. Find FRET plasmids here. Split EGFP: An alternative to FRET, split EGFP has also been... uses of GFP. Förster resonance energy transfer (FRET): This is used to study the interactions between...concentrations and pH, using a range of strategies such as FRET, calmodulin, and others. Review Addgene's collection...

... tips from experts on FRET techniques. By combining standard donor/acceptor FRET know-how with aptamers...Aptabeacons Fluorescence resonance energy transfer (FRET) technology has been studied and used in assays ...Additional Resources on the Addgene Blog Tips for Using FRET in Your Experiments Learn more about GFP Controlling...

...evaluated set of values.  However, to evaluate the interpretation of the experiment would require access to each...no one is suggesting all data be stored in a retrievable format, we too rarely preserve “raw” data of ...journal repository, they are likely too large to be retrieved by others. Other new data initiatives are also...addition to enhancing veracity of collections and interpretation of data, publishing complete data, rather than...

...Filonov et al., 2013). FRET biosensors were developed based on miRFP760-miRFP720 FRET pair, including Rac1...indicator (Shemetov et al., 2021). In addition to FRET, intensiometric calcium indicators NIR-GECOs were...optogenetics of Rho GTPases enabled by near-infrared FRET. Nat Chem Biol 14, 591-600 (2018). Shemetov, A.A...

...Evaluating Efficiencies of Dual AAV Approaches for Retinal Targeting. Front Neurosci 11: . https://doi.org...(2013) Effective delivery of large genes to the retina by dual AAV vectors. EMBO Mol Med 6:194–211 . https...truncated proteins are safe and effective in the retina of a mouse model of Stargardt disease. Hum Mol ...first time AAV user Learn about how AAV is used in retinal gene therapy Learn about the parts of an AAV transfer...

...For cloning into and storage of lentiviral and retroviral vectors or cloning or repeated sequences with...cloning and storage of unstable DNA and lentiviral/retroviral vectors. Used by Addgene for pooled library amplification...

...or PNS transduction, cell type specificity, or retrograde transport in neurons. (b) Genetic cargo can be...typically administered in the lateral tail vein or retro-orbital sinus. IT injections are administered in... Chen et al., 2022 PAL2 CNS: cerebrum; neuroretina Macaques N/A Stanton et al., 2023 AAV-MDV1A...could be used to target endothelial cells in the retina and spinal cord (Krolak et al., 2022). Systemic...macaques. It also effectively transduces the neuroretina. PAL2 detargets the liver and has increased DRG...Zheng, Q., Kozareva, V., Beddow, T., Tobey, I. G., Pacouret, S., Chen, A. T., Chan, Y. A., Ryvkin, D., Gu,...

...remember that some serotypes can be transported retrogradely and/or anterogradely along the axon. Others,...your gene and could lead to toxicity or trouble interpreting your results, while a low transduction efficiency...Puschnik, A. S., Nagamine, C. M., Cheng, F., Qiu, J., Carette, J. E., & Vandenberghe, L. H. (2018). An Alternate...Introduction to AAV Viral Vectors 101: The AAV Retrograde Serotype Viral Vectors 101: Viral Applications...

...to not have any effect on each other, they have discrete effects on the dependent variable, and those variables...others. Each test has different likelihoods of returning false positives/negatives, but they all generally...numerical value relative to quantiles derived from a theoretical distribution. If your data is normally distributed...130 (2012). 10.1097/SMJ.0b013e3182498ad5 Curran-Everett, Douglas. Explorations in statistics: statistical...

...Working on the Eye 9:05-10:16 : Cell Targeting in Retinal Gene Therapies 10:17-14:34 : Progress in Treating...Treating Macular Degeneration and Other Retinopathies 14:39-25:59 : Strategies to Make Gene Therapies More... retina...

...to tell us the story of his incredibly popular retroviral vector pBABE-puro. Read on to learn more about...well as Dusty Miller’s lab, we set out to derive retroviral vectors that would yield the highest titers whilst...recombination that would lead to replication competent retrovirus.  When we arrived at the final vectors (including...

... user, instead, don’t fret; we have backgrounds for you as well! If you use FRET, you might be interested...Fluorescent Proteins blog topic to learn more about FRET and other techniques using FPs! Otherwise, check...

...find this hard so get used to it by making it a concrete goal to say “thank you, job well done” to one ...not the person - intent is usually good. Don’t pretend something went well when it didn’t… but don’t “...compare with your own perception? Tell me your interpretation of my feedback to you. What are We Learning...

...increased yields in adenoviral (Coroadinha, 2006) and retroviral production systems (Shen, 2011). To our knowledge...also improved yields of the AAV2 derivative, rAAV2-retro, but had no effect on AAV5 yields. We further show...Effect of osmotic pressure on the production of retroviral vectors: Enhancement in vector stability." Biotechnology...