...indicates a virus containing the genome of serotype 2 packaged in the capsid from serotype 5. Use of these...neurons that are not efficiently transduced by AAV2/2, and is distributed more widely in the brain, indicating... pseudotyped with viral capsids from serotypes 1, 2, and 5 display differential efficiency and cell tropism...PJ, Mandel RJ, Muzyczka N. Mol. Ther. 2004 Aug;10(2):302-17. PubMed . From virus evolution to vector revolution... Science Guides AAV Guide Adeno-associated Virus (AAV) Guide Components...Viral serotypes and pseudotyping/gene therapy: Designer gene delivery vectors: molecular engineering and...

...//doi.org/10.1002/1096-8628(200022)97:2<147::aid-ajmg6>3.0.co;2-o Baker SA, Chen L, Wilkins AD, Yu P, ...Caused by a mutation in the methyl-CpG binding protein 2 (MECP2) gene, Rett Syndrome is a rare, progressive...in transcriptional repression, activity dependent de-repression, chromatin remodeling, gene activation...X-linked MECP2, encoding methyl-CpG-binding protein 2. Nat Genet 23:185–188 . https://doi.org/10.1038/13810... (2000) Rett syndrome: Methyl-CpG-binding protein 2 mutations and phenotype-genotype correlations. Am ...Tillotson R, Selfridge J, Koerner MV, Gadalla KKE, Guy J, De Sousa D, Hector RD, Cobb SR, Bird A (2017) Radically...that MECP2 binds to methylated CpG dinucleotides and CAC trinucleotides during neuronal maturation (Tillotson...

...is an encyclopedia of Class 2 CRISPR systems with wiki entries describing enzyme activity, experimental...Resources CRISPR Guide Viral Preps Protocols gRNA Design Tools CRISPR Blog Posts Two classes of base editors...protein (Mu- GAM). While many base editors are designed to work in a very narrow window proximal to the...

...postdoctoral researcher at Instituto de Investigación Biomédica de Salamanca (IBSAL) and one of the coordinators...mammal called alpaca could neutralize the SARS-CoV-2 virus. But while the early results were promising,...receiving dozens of requests every day.    Figure 2: When a researcher needs a collaborator for a specific...Crowdfight will: 1) Find an expert with the right skill; 2) Confirm the expert’s suitability and availability... illustrates the power of collaboration, which despite the competitiveness of cutting-edge research, is...physics. Even non-scientists, such as lawyers, designers and other professionals, joined the platform; ...

...thanks to our guest blogger, Harshana S De Silva Feelixge. Harshana S De Silva Feelixge is a researcher whose... post was contributed by guest blogger Harshana S De Silva Feelixge. Gene therapy technologies hold ...AAVs in the clinic Thus far, AAV vectors AAV1, AAV-2, AAV1-AAV2 hybrids, AAV-6, AAV-7, AAV-8, AAV-9 and...neutralizing antibodies against some AAV types, AAV1, 2, 3 and 5 are thought to be as high as 70% among the... 10.24 (2003): 1999-2004. PubMed PMID: 14566358.  2. Hirsch, M. L., et al. "Self-complementary AAV mediates...form, translating gene therapies from conceptual design to clinical trials involve identifying a therapeutic...

...Cre-ERT2 CAG Mammalian Heller 112622 pVHC_PGKneoLox2DTA.2 Venus, Cre-ERT2, targeting vector with MCS for homology...homology arms Mammalian Heller 112623 pEHC_PGKneoLox2DTA.2 Emerald, Cre-ERT2, targeting vector with MCS for homology... arms Mammalian Heller 112624 pmTHC_PGKneoLox2DTA.2 TFP, Cre-ERT2, targeting vector with MCS for homology...arms Mammalian Heller 112625 ptdTHC_PGKneoLox2DTA.2 tdTomato, Cre-ERT2, targeting vector with MCS for ...25;1504(4):467-86. doi:10.1016/0022-2836(81)90375-2. PubMed . Ventura A, Meissner A, Jaenisch R, Jacks...efficiency, selecting desired clones, and making transgenic mice. ID Plasmid/Article Description Expression System...Cre expression plasmids. Addgene's viral service provides high-quality and high-titer viral preparations...

...SARS-CoV-2 spike pseudotyping, visit the SARS-CoV-2 Pseudotyped Virus collection page. SARS-CoV-2 detection...of SARS-CoV-2 entry and become upregulated during viral infection.  Expression of SARS-CoV-2 viral proteins...McLellan JS (2020) Structure-based design of prefusion-stabilized SARS-CoV-2 spikes. Science 369:1501–1505....labs in over 75 countries. Figure 1: SARS-CoV-2 proteins and other components. Image from Maya Kostman... protein, variants, and pseudotyping The SARS-CoV-2 virus depends on its spike protein to bind ACE2, the...of infection and how to interfere with it in a BSL-2 lab instead of a BSL-3 lab. Jesse Bloom’s lab published...reagents for pseudotyping lentivirus with the SARS-CoV-2 spike protein. They deposited the plasmids used for...

...none S. pyogenes de Bono sgRNA with rpr-1 promoter 48961 Worm EcoRI none S. pyogenes de Bono pMB60 47941...is an encyclopedia of Class 2 CRISPR systems with wiki entries describing enzyme activity, experimental...BclI-SwaI none S. pyogenes URA3 Wyrick pCRISPomyces-2 61737 Bacteria BbsI yes, cut S. pyogenes Apramycin... S. pyogenes Zhang gRNA Cloning Vector Bbs I ver. 2 85586 Mammalian BbsI None S. pyogenes H Fujii pX601...pyogenes LEU2 Lu pDestTol2CG2-U6:gRNA 63156 Zebrafish BseRI none S. pyogenes Zon pDestTol2pA2-U6:gRNA 63157...module vectors, including three designed for dicots and three designed for monocots. Using these gRNA ...Resources CRISPR Guide Viral Preps Protocols gRNA Design Tools CRISPR Blog Posts CRISPR requires that you...

...goals when designing and testing your AAV.  Figure 2: Comparison of AAV serotypes 1, 2, 5, 6, 8,...) your experimental conditions and measurements; (2) the cells you’re targeting; (3) how much expression...as your titer changes, so may your needed volume (de Backer et al., 2010).  Second, high efficiency transduction... Tropism and Axonal Transport of AAV Serotypes 1, 2, 5, 6, 8 and 9 in the Mouse Brain. PLOS ONE, 8(9),...–10. https://doi.org/10.1371/journal.pone.0106472 de Backer, M. W. A., Brans, M. A. D., Luijendijk, M.... Comparison of four promoters. Gene Therapy, 28(1–2), 56–74. https://doi.org/10.1038/s41434-020-0169-1...analyses of adeno-associated viral vector serotypes 1, 2, 5, 8 and 9 in marmoset, mouse and macaque cerebral...

...backbone which is described by Durocher et al. Nucleic Acids Research 2002 Jan 15;30(2):E9 A list of commonly-used...showing how AVEXIS works with a detailed protocol description: Avidity based Extracellular Interaction Screening...

...nucleases. Genome Res. 2010 Jan;20(1):81-89. PubMed. 2. Bauer DE, Kamran SC, Lessard S, Xu J, Fujiwara Y, Lin ...with TALENs, in collaboration with the Porteus lab [2]. With the advent of CRISPR/Cas9, we began to explore...11;342(6155):253-257. PubMed. 3. Canver MC, Bauer DE, Dass A, Yien YY, Chung J, Masuda T, et al. Characterization...and Bauer lab's related paper describes their methods for CRISPR design, cloning, and delivery, as well...

...on opposite sides of the world shows that science is truly a global endeavor. Case study 2: Plasmids Another...repository provides. Repositories around the world facilitate the rapid pace of SARS-CoV-2 research by...fast access to the tools required to study SARS-CoV-2, but with many academic research labs closed during...biobanks, are crucial for the advancement of SARS-CoV-2 research because they provide rapid access to validated...Many of the reagents used to investigate SARS-CoV-2 were developed for broad use and are now important...susceptibility of cells from different organisms to SARS-CoV-2 (Chu et al., 2020). An additional cell line used in...applications but have proven useful to the study of SARS-CoV-2 are the lentiviral production and packaging vectors...

...suggestions, like alternative word suggestions. De-jargonizer The De-jargonizer can identify jargon in your writing...episode 2 "Organizing Literature" : I found out about Zotero here Dear Grad Student podcast episode 2 "Essential...This paper talks more about the development of the De-jargonizer.   Academic Phrasebank  The Academic Phrasebank... of the suggestions Hemmngway makes (it wasn’t designed for technical writing), it’s great at flagging... Just enter the DOI of a paper and select your desired formatting style. This tool is great for generating...library in Microsoft Word. Mendeley has a web and desktop version that works with Windows, Mac, and Linux...easier to work with collaborators. Zotero is a desktop based program that’s compatible with Mac, Windows...

... and G-protein is delivered to the starter cell. (2) The starter cell now expresses TVA and G-protein....Vectors for Monosynaptic Neuronal Tracing ID Name Description Serotype PI 52473 pAAV-synP-FLEX-splitTVA-EGFP-B19G...

...Information Featured Collections COVID-19 SARS-COV-2, ACE2, and CRISPR tools Fluorescent Proteins from ...materials and intellectual property rights, and provides liability protection for material providers. Addgene...

...upstream of an opsin gene (e.g., channelrhodopsin-2, ChR2) fused to a reporter gene (e.g., mCherry) to...the correct orientation to be transcribed (Figure 2) (Sharma and Zhu, 2014). How does FLEx enable strong... rather than aversive stimuli. In a recent paper, de Jong and colleagues used rAAV FLEx-based vectors ...brain regions associated with motivated behaviors (de Jong et al., 2019). FLEx vectors represent, therefore...developmental biology at the École Normale Supérieure de Lyon. Her love for educational science led her to... Abdallah, Khaled, et al. "Adeno-associated virus 2/9 delivery of Cre recombinase in mouse primary afferents...Deniz, et al. "A FLEX switch targets Channelrhodopsin-2 to multiple cell types for imaging and long-range ...

...Cas9 plasmid Bullock and Port pCFD2-dU6:2gRNA dU6:2 BbsI Injection or in vitro transcription Virmilion...Protocols gRNA Design Tools CRISPR Blog Posts The following CRISPR plasmids have been designed for use in ... to a genomic DNA locus and encodes a primer binding site and the desired edits on an RT template. Plasmid...CRISPR plasmids that have been designed for use in Drosophila and other insects. CRISPR... can increase transcription of a specific gene. Design your gRNA sequence to direct the dCas9-activator...

....9 (parental) 46 +/- 0.7% AAV.PHP.S 82 +/- 2% Table 2: Transduction efficiency of AAV.9 versus AAV.PHP.S...figure 2) to allow for efficient IV delivery of virus to the CNS or PNS (figure32, tables 1 & 2). These...variants for widespread gene transfer to the adult brain. Nature biotechnology. 2016;34(2):204-209. PubMed...because: 1) their genomes are easy to manipulate, 2) they have long-term expression; and 3) they have ... On average, AAV.PHP.eB and AAV.PHP.S had 1.5- to 2-fold higher GFP intensity compared to the strains ...stained for a neuronal marker). Check out tables 1 and 2 below for a summary of the quantified transduction...cells are labeled with their own unique color; and 2) low levels of labeling so that a cell’s projections...

...applications. LuxSit-i : An artificial luciferase desined de novo using deep-learning approaches. Split-Luciferase...fusing NanoLuc® and Venus fluorophore to Troponin C Ca 2+ binding domain. Nano-lanterns , Enhanced Nano-lanterns...transfection Feng Zhang 21471 pLenti PGK V5-LUC Neo (w623-2) Firefly PGK Lentiviral expression of firefly luciferase...to that of the fluorophore. Conformationally destabilized Firefly luciferase mutants : A set of three ...three sensors that are increasingly structurally destabilized versions of Firefly luciferase which serves ...dimerization , endosomal escape of cell-penetrating peptides , and more. nLuc/cLuc for split Firefly Luciferase...mammalian systems. ID Plasmid Luciferase Type(s) Description PI 16539 pBV-Luc Firefly Insertion of 5' promoter...

...is an encyclopedia of Class 2 CRISPR systems with wiki entries describing enzyme activity, experimental...Sequences Glossary Publications CRISPR Overview Class 2 C lustered R egularly I nterspaced S hort P alindromic...systems enable researchers to target anywhere from 2 to 7 genetic loci by cloning multiple gRNAs into a... is included on the gRNA-containing plasmid, or a 2-plasmid system in which Cas9 must be delivered separately...library (panel E ). Remember - if you are using a 2-vector system, you will need to transduce cells that...either (1) a lack of gRNA and/or Cas9 expression or (2) a lack of efficient target cleavage in cells expressing...off-target effects by using a single Cas9 nickase and 2 different gRNAs, which bind in close proximity on ...