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TypeBlog PostPublishedSept. 6, 2022, 1:15 p.m....Kostick, K., Torgerson, L., Muñoz, K. A., Hsu, R., Kalwani, L., Sierra-Mercado, D., Robinson, J. O., Outram... rat CNS: AAV PHP.EB vector dose–response and comparison to AAV PHP.B. Gene Therapy, 25(5), 392–400. https...00981-0 Wright, J. F., Le, T., Prado, J., Bahr-Davidson, J., Smith, P. H., Zhen, Z., Sommer, J. M., Pierce... Jason Nasse...
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Simplify Cloning with in vivo Assembly
TypeBlog PostPublishedOct. 18, 2018, 12:37 p.m.... Jake Watson and Javier García-Nafría from the MRC Laboratory of Molecular Biology. Jake Watson is a postdoctoral...This post was contributed by Jake Watson and Javier García-Nafría from the MRC Laboratory of Molecular...receptor function. Follow them on Twitter @jakefwatson and @JGarciaNafria. References 1. Jones, Douglas...PMCID: PMC331480. 3. García-Nafría, Javier, Jake F. Watson, and Ingo H. Greger. "IVA cloning: a single-tube... -
New Optimized Genome-wide CRISPRko, CRISPRi, and CRISPRa Libraries
TypeBlog PostPublishedOct. 4, 2018, 12:44 p.m....libraries that do just that (Doench et al., 2016, Sanson et al., 2018). These libraries utilize optimized...additional transcription factors to the targeted region (Sanson et al., 2018) . This tracrRNA was combined with...selected per gene and divided into two sets, A and B. Sanson et al. demonstrated that the Calabrese library ... more hits than the SAM library approach in a comparison study screening for vemurafenib resistance in...: 25494202. PubMed Centra PMCID: MC4420636. 5. Sanson, Kendall R., et al. "Up, down, and out: next generation... -
CRISPR 101: Cytosine and Adenine Base Editors
TypeBlog PostPublishedFeb. 13, 2025, 2:15 p.m....favorably to other methods. In a head-to-head comparison with Cas9, Cas9 modified 9/12 known off-targets... C., Rees, H. A., Packer, M. S., Badran, A. H., Bryson, D. I., & Liu, D. R. (2017). Programmable base ...doi.org/10.1038/nbt.3803 Koblan, L. W., Doman, J. L., Wilson, C., Levy, J. M., Tay, T., Newby, G. A., Maianti... Bohnuud, T., Fernandez, T. P., Gehrke, J. M., Grayson, P., Lee, K. D., Ortega, M. A., Sawyer, C., Schwaegerle..., Lapinaite, A., Newby, G. A., Thuronyi, B. W., Wilson, C., Koblan, L. W., Zeng, J., Bauer, D. E., Doudna...Levy, J. M., Yeh, W.-H., Zheng, C., Newby, G. A., Wilson, C., Bhaumik, M., Shubina-Oleinik, O., Holt, J.... -
How to Be an Excellent Trainee
TypeBlog PostPublishedSept. 26, 2023, 1:15 p.m....advice, open doors for you, and teach you valuable lessons along the way. We think it’s so important that ... technician, or a graduate student. This is the person you report or talk to on a daily basis who will...scared to ask for some! You can email or ask in-person “Are there any papers you think would be useful...these things will help you get more out of the in-person training component. Documenting during training...take on trainees one day when you are the senior personnel! Happy training! Resources Resources on the Addgene... -
Twenty Years of Sharing: Addgene's Viral Vector Service
TypeBlog PostPublishedJuly 30, 2024, 1:15 p.m....service in 2016. We realized we could apply the lessons learned from our plasmid lab to develop a reliable...available. This prioritization happens for good reasons. We began offering our viral vector service because...our Viral Vectors 101 eBook and blog posts. The lessons learned from this process have shaped and supported...fnana.2019.00093 McDade, J. R., Waxmonsky, N. C., Swanson, L. E., & Fan, M. (2016). Practical Considerations... Rachel Leeson... -
Neuronal labeling with Spaghetti Monster
TypeBlog PostPublishedAug. 14, 2018, 12:42 p.m..... 4. Shaner, Nathan C., George H. Patterson, and Michael W. Davidson. "Advances in fluorescent protein... structure resembles images of the deity (see comparison image above). The church of the Flying Spaghetti...schools and was started in 2005 in Kansas by Bobby Henderson, a physics graduate from Oregon State University...New York. pp 719. 3.Rizzo, Mark A., Michael W. Davidson, and David W. Piston. "Fluorescent protein tracking... -
Transferable Skills Guide: External Collaborations
TypeBlog PostPublishedApril 19, 2018, 1:25 p.m....know your collaborators from other institutions personally on conference calls and make sure to maintain...collaborating with people outside my lab’s department, I personally felt that I had to overcome some hesitation to...the hard part: maintaining these relationships. Personally, I find this to be the most challenging and time-intensive...maintain a relationship - many of them involve personal meetings and common interest so naturally this...sharing. LinkedIn or ResearchGate allow you to send personal updates as messages and keep you up-to-date on... -
AAV Titers: Where Do They Come From And What Do They Mean?
TypeBlog PostPublishedNov. 15, 2017, 3 p.m....are addressed with ddPCR technology (Hindson et al., 2011, Hindson et al., 2013), which has been shown ...2011): 8604-8610.4. Hindson CM, Chevillet JR, Briggs HA, Gallichotte EN, Ruf IK, Hindson BJ, Vessella RL,... therapy 6.7 (1999). PubMed PMID: 10455443. 3. Hindson, Benjamin J., et al. "High-throughput droplet digital... -
Rett Syndrome: A History of Research and Therapeutic Outlooks
TypeBlog PostPublishedNov. 19, 2020, 2:15 p.m....CAC trinucleotides during neuronal maturation (Tillotson & Bird, 2019) and is a global transcriptional ...rescue the neurological defects of Rett syndrome (Tillotson et al., 2017). Advancing gene therapies for Rett...Supplement/S51.002.abstract Sinnamon JR, Kim SY, Fisk JR, Song Z, Nakai H, Jeng S, McWeeney SK, Mandel G (2020)... . https://doi.org/10.1016/j.omtm.2017.04.006 Tillotson R, Selfridge J, Koerner MV, Gadalla KKE, Guy J...398–401 . https://doi.org/10.1038/nature24058 Tillotson R, Bird A (2020) The Molecular Basis of MeCP2 ... -
Plasmids 101: Codon usage bias
TypeBlog PostPublishedSept. 27, 2018, 1:09 p.m....organisms have different codon usage biases? The reasons for varied codon preferences among organisms aren...aren’t completely understood, but some possible reasons include: Metabolic pressures - it takes cellular...recognize codons found in highly expressed genes (Emilsson and Kurland, 1990). Controlling gene expression...15678157. PubMed Central PMCID: PMC1299251. 3. Emilsson, Valur, and Charles G. Kurland. "Growth rate dependence... 2265611. PubMed Central PMCID: PMC552224. 4. Gustafsson, Claes, Sridhar Govindarajan, and Jeremy Minshull... -
CRISPR Activation: A Practical Guide
TypeBlog PostPublishedAug. 18, 2016, 2:30 p.m....Pinnelli M, Ansari R, Harper S, Jackson DA, McRae R, Pooley R, Wilkinson P, van der Meer D, Dow D, Buser-Doepner...of TSS for best results but up to 400 bp works reasonably well. Best of luck on your experiments! Many...Housden BE, Perrimon N, Collins JJ, Church G (2016) Comparison of Cas9 activators in multiple species. Nat Methods...doi.org/10.1038/nmeth.3871 Gilbert LA, Horlbeck MA, Adamson B, Villalta JE, Chen Y, Whitehead EH, Guimaraes... -
Antibodies 101: Single Chain Fragment Variables (scFvs)
TypeBlog PostPublishedJune 3, 2021, 1:15 p.m....10.3390/antib8020028 Bird R, Hardman K, Jacobson J, Johnson S, Kaufman B, Lee S, Lee T, Pope S, Riordan... (Sandomenico et al., 2020). Figure 1: Comparison between the IgG antibody and scFv. What...doi.org/10.1371/journal.pone.0189964 Huston JS, Levinson D, Mudgett-Hunter M, Tai MS, Novotny J, Margolies... made by molecular evolution 1 1Edited by I. A. Wilson. Journal of Molecular Biology 275:245–253 . https... -
Five Popular Model Organisms
TypeBlog PostPublishedApril 11, 2019, 12:39 p.m....cells are great model organism not only for the reasons above but because they actually share many biological... the study of neurological diseases, such as Parkinson’s disease (PD). PD is primarily characterized by...pancreas and produce different levels of insulin (Hesselson et al., 2009). The worm (Caenorhabditis elegans...nature391.6669 (1998): 806. PubMed PMID: 9486653. Hesselson, Daniel, et al. "Distinct populations of quiescent...From the baker to the bedside: yeast models of Parkinson's disease." Microbial cell 2.8 (2015): 262. PubMed... -
Viral Vectors 101: AAV Variables That Matter
TypeBlog PostPublishedJune 1, 2023, 1:15 p.m....designing and testing your AAV. Figure 2: Comparison of AAV serotypes 1, 2, 5, 6, 8, and 9 in two ...Taschenberger et al., 2017). Figure 3: Comparison of two astrocyte selective promoters demonstrating...Sortwell, C. E., Hauswirth, W. W., Boye, S. E., & Manfredsson, F. P. (2017). Rationally Engineered AAV Capsids...vector-mediated transduction of the corticospinal tract: Comparison of four promoters. Gene Therapy, 28(1–2), 56–... Jason Nasse... -
Plasmids 101: Dimers and Multimers
TypeBlog PostPublishedJan. 10, 2023, 2:15 p.m....PMC2693335. Williams JA, Luke J, Langtry S, Anderson S, Hodgson CP, Carnes AE. Generic plasmid DNA production...and at high copy numbers (Williams, Carnes, and Hodgson, 2009). More research is ongoing to determine plasmid...90057-7. PMID: 2544913. Cook KS, Luo J, Guttman A, Thompson L. Vaccine Plasmid Topology Monitoring by Capillary...90060-6. PMID: 6323019. Williams JA, Carnes AE, Hodgson CP. Plasmid DNA vaccine vector design: impact on... -
An “elegans” Approach to Better CRISPR/Cas9 Editing Efficiency
TypeBlog PostPublishedDec. 17, 2020, 1:30 p.m....2013), or rescue of mutant phenotypes (unc-119) (Dickinson et al., 2013). These approaches allowed effective...the development of the self-excising cassette (Dickinson et al., 2015) made this step as simple as heat-shocking...e193–e193 . https://doi.org/10.1093/nar/gkt805 Dickinson DJ, Pani AM, Heppert JK, Higgins CD, Goldstein...sequences https://doi.org/10.17912/W2237D Farboud B, Severson AF, Meyer BJ (2018) Strategies for Efficient Genome...Krishnamurthy PM, Gneid H, Edraki A, Watts JK, Sontheimer EJ, Mello CC (2018) 5′ Modifications Improve ... -
Fluorescent Proteins 101: Photoactivatable Fluorescent Proteins
TypeBlog PostPublishedApril 25, 2017, 2:30 p.m....many stages of synthesis and degradation. For this reason it is virtually impossible to determine specific...parameter to consider is the contrast ratio- a comparison of PA-FP brightness before and after photoactivation...extraordinary molecular detail. References 1.Patterson, George H., and Jennifer Lippincott-Schwartz. ..., Samuel T., Thanu PK Girirajan, and Michael D. Mason. "Ultra-high resolution imaging by fluorescence ...Photoactivatable FPs in Empty Backbones in the Michael Davidson Collection Find Photoactivatable FPs for Single... -
Editor's Choice, September 2016
TypeBlog PostPublishedOct. 7, 2016, 4 p.m....contributed by Kurt Swanson, husband of Addgene Director of Biology, Lianna Swanson, and researcher at ...in food products). I picked this post for a few reasons: 1) I think phage are super cool and have considered...proteins that could be useful for a wide variety of reasons. 3) I would love to see more posts like this on... -
28 Hot Plasmid Technologies from 2015
TypeBlog PostPublishedDec. 23, 2015, 3:30 p.m....use Bioluminescence Resonance Energy Transfer (BRET) were developed by the Johnson Lab . Characteristics...CRISPR labeling chromosomal loci in genome Thoru Pederson’s lab at the University of Massachusetts Medical...lab further modified sdChr- a blue-shifted channelrhodpson from freshwater green alga identified by Ed...Nano-lanterns rely on the principle of bioluminescence resonance energy transfer (BRET), whereby photons emitted...-tagged human ORF clones to allow testing and comparison of different cell-free expression systems; the...understanding complex gene regulation (Bert et al 2000, Johnson et al 2004). So if you are looking for a luciferase... et al. Plasmid. 2000. PubMed PMID: 10964627 Johnson, et al. Mol Cell Biol. 2004. PubMed PMID: 15340054...