We narrowed to 185 results for: INV-E
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TypeBlog Post...PubMed Central PMCID: PMC3712628. 5. Dahlman, James E., et al. "Orthogonal gene knockout and activation ...and of modulating gene expression makes it an invaluable tool in the interrogation of gene function. However...
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Anatomy of a Plasmid Page at Addgene
TypeBlog Post...strain. Most Addgene plasmids are maintained in the E. coli lab strain DH5a, but others may require strains...? Depositing lab: The name of the Principal Investigator (PI) in whose lab the construct was made. The... -
Interview: Nicola Patron on Plant Synthetic Biology, MoClo, and More
TypeBlog Post...standardized assembly of multigene constructs. Weber E, Engler C, Gruetzner R, Werner S, Marillonnet S. PLoS...that will provide a similar resource for those involved in monocot engineering projects. The MoClo tools... -
CRISPR 101: Ribonucleoprotein (RNP) Delivery
TypeBlog Post...approach is to express and purify His-tagged Cas9 from E. coli using a plasmid such as pET-28b-Cas9-His from...Chen et al. 2016). For CRISPR experiments that involve HDR, electroporation can be combined with cell-... -
Sonic Hedgehog? Sleeping Beauty? Learn About the Genes Behind Addgene’s Conference Room Names
TypeBlog Post...2003): 563-573. PubMed PMID:12490562. Karess, Roger E., et al. "The regulatory light chain of nonmuscle ...factors to repress the expression of genes primarily involved in development. These genes are known to play ... a comic book his daughter was reading. SHH is involved in the hedgehog signalling pathway, which is a...resemble spaghetti. We have two plasmids in our inventory from the lab of Eric Wieschaus that express sqh...Azplazu and Frasch, 1993). Tinman is specifically involved in the formation of the heart amongst other things...Transcriptional activation in Drosophila spermatogenesis involves the mutually dependent function of aly and a novel... -
Production of Virus in Insect Versus Mammalian Cells
TypeBlog Post...–871. https://doi.org/10.1096/fj.01-0995fje. Tan, E., Chin, C. S. H., Lim, Z. F. S., & Ng, S. K. (2021... methods add an additional viral passage. This involves taking your first round of virus, called P1 (or... -
Don’t FRET: Bimolecular Fluorescence Complementation Makes Visualizing Protein-Protein Interactions Easy
TypeBlog Post...grid. References Bischof J, Duffraisse M, Furger E, Ajuria L, Giraud G, Vanderperre S, Paul R, Björklund... to be successful. A BiFC experiment generally involves three steps: fusion protein construction, cellular... -
Beyond the Blood Brain Barrier: PHP.B Enters the Central Nervous System Via the LY6A Receptor
TypeBlog Post...PubMed Central PMCID: PMC5529245. Deverman, Benjamin E., et al. "Cre-dependent selection yields AAV variants...Ken Y., et al. "Engineered AAVs for efficient noninvasive gene delivery to the central and peripheral nervous... -
New Tools Enable CRISPRa for Neuroscience Applications
TypeBlog Post...PubMed Central PMCID: PMC6214761. Savell, Katherine E., et al. "A neuron-optimized CRISPR/dCas9 activation...singled out two Bdnf isoforms (I and IV) for investigation. Co-transduction of primary rat neurons with... -
Hot Plasmids - August 2020
TypeBlog Post...Nanobodies are easily expressed in the cytosol of E. coli, however the nanobody proteins tend to get stuck... at small and large scale. This new technique involves selection in the cytosol (instead of on the yeast... -
The Fluorescent Vegetables in Aptamer Soup
TypeBlog Post...Nilaratanakul, Voraphoj, Debra A. Hauer, and Diane E. Griffin. "Development and characterization of sindbis...2013). Since then, Spinach2 has been used to investigate intracellular viral DNA dynamics in neuronal ...developed into a split aptamer that can be used to investigate RNA assembly. The split aptamer is composed of... blog Learn more about aptamer fluorophores Investigate other fluorescent biosensors Find other uses ... -
Adeno-associated Viruses (AAVs) for Genome Editing
TypeBlog Post...219. PubMed PMID: 27280971. 10. Epstein, Benjamin E., and David V. Schaffer. "Combining engineered nucleases...delivered through transfection of naked DNA. Other investigators who had been working to develop high efficiency..., which is particularly well suited for strand invasion, a key step in the homologous recombination pathway... -
Antibodies 101: Introduction to Immunofluorescence
TypeBlog Post...Bandrowski A, Carr S, Edwards A, Ellenberg J, Lundberg E, Rimm DL, Rodriguez H, Hiltke T, Snyder M, Yamamoto...sample type. Sample Preparation This step can involve multiple smaller steps that all help to ensure ... Similar to Sample Preparation, this step can involve multiple smaller steps, but the aim is to preserve... -
Rett Syndrome: A History of Research and Therapeutic Outlooks
TypeBlog Post...Rinaldi F, Wein N, Meyer KC, Solano S, Nguyen K , Lang E, Kaspar AA, Foust KD, Thomsen G, Fugere M, Kaspar ...). Beam Therapeutics, co-founded by Harvard Investigators and Addgene depositors David Liu, PhD, Keith... -
Plasmids 101: Plasmid Incompatibility
TypeBlog Post...Pubmed PMID: 28245276 Liu X, Wang D, Wang H, Feng E, Zhu L, Wang H (2012) Curing of Plasmid pXO1 from ... plasmids encode a negative regulation system involving antisense RNAs or iterons that inhibits replication... -
Tips and Tricks for Using Golden Gate Modular Cloning (MoClo)
TypeBlog Post...plate after each transformation and/or use different E. coli strains with higher transformation efficiency...of the developers of the MoClo system, are an invaluable resource for learning about MoClo in an in-depth... -
Bacterial Expression Systems
TypeCollection...Gate modular cloning (MoClo) for use in E. coli . Addgene Blog E. Coli Strains for Protein Expression Protein... to purify it. Specific bacterial strains such as E. coli BL21(DE3) are used as biofactories to express...cytoplasmic and periplasmic protein expression in E. coli , Bacillus subtilis , and Pichia pastoris . ... (reconstructed) BiFC Dan Mulvihill 39866 pWA PAS-E BAD K-GAFm iRFP (reconstructed) BiFC Vladislav Verkhusha...reliably drive expression of a gene of interest in E. coli . The Marionette Sensor Collection (Addgene ...twelve genes using small-molecule inducers in the same E. coli strain. Addgene Blog Inducible Promoters Repressible...Promoters Synthetic Photobiology: Optogenetics for E. coli Return to top Reporter Plasmids Reporter plasmids... -
Building Global Connections with the International Mentorship Program USA-EUROPE
TypeBlog Post... conferences and had real-time correspondence via e-mail throughout the entire year. In addition to their...base of former mentees, we encourage them to get involved and give back by becoming mentors to new students... -
How to Design Your gRNA for CRISPR Genome Editing
TypeBlog Post...Younger ST, Zhivich V, Najgebauer H, Allen F, Gonçalves E, Shepherd R, Doench JG, Yusa K, Vazquez F, Parts L...to perform functional screens to identify genes involved in a phenotype of interest. This blog post will... is a very low-efficiency process, and usually involves the need for single cell cloning and subsequent... -
Custom CRISPR Screens & the Green Listed Software
TypeBlog Post...PubMed Central PMCID: PMC4744125. 3. Sanjana, Neville E., Ophir Shalem, and Feng Zhang. "Improved vectors ...targeting each of the 1000 genes you’d like to investigate in your next CRISPR screen. Luckily, the Green... unbiased discovery. With these screens, the investigator generates a cell population where all genes ...consequence, which genes are good candidates for being involved in the studied phenotype. For robustness of a ... generated knockout mice for (all of the genes involved as I recall). The lab then used these knockout...sequenced to identify which signaling molecules were involved in the phenotype. Surface proteins My group works... These types of interactions to a large extent involve receptors expressed on the cell surface. Thus, ... -
CRISPR History and Development for Genome Engineering
TypeCollection...Repeat) sequences were initially discovered in the E. coli genome in 1987, but their function as a safeguard...Mammalian systems (Human, mouse, and rat) Bacteria ( E. coli , Streptococcus , Streptomyces , and others)...Chylinski K, Fonfara I, Hauer M, Doudna JA, Charpentier E. 2012. A programmable dual-RNA-guided DNA endonuclease... Saunders SJ, Barrangou R, Brouns SJ, Charpentier E, Haft DH, Horvath P, Moineau S, Mojica FJ, Terns RM...9. PMID: 26355004 Shalem O, Sanjana NE, Hartenian E, Shi X, Scott DA, Mikkelson T, Heckl D, Ebert BL, ...as a powerful defensive strategy against viral invaders, and this system is proving to be just as useful...CRISPR-associated (Cas) genes to not only store a record of invading phages but also to destroy the phages upon re-... -
A Guide to Getting Started in Undergrad Research
TypeBlog Post...https://doi.org/10.1126/science.351.6276.994 Moore E (2021) One mentor isn’t enough. Here’s how I built...postdoc” is, and that “PI” stood for principal investigator. In basic science (as opposed to translational...in a lab. Position Common tasks Principle investigator (PI) Runs the lab Writes a lot of grants Varying...prep media, etc. "Research assistants" are more involved in the science, experimental design, and execution...genuinely excites them, and reach out to the PI to get involved. It not only demonstrates initiative and drive... -
FLEx Technology and Optogenetics: Flipping the switch on gene expression with high spatial and temporal resolution
TypeBlog Post... PubMed Central PMCID: PMC4100247. Taylor, Norman E., et al. "Optogenetic activation of dopamine neurons...and optogenetics meet, allowing scientists to investigate the fundamental basis of both brain function ... flip-excision) switch (aka DIO (Double-floxed Inverse Orientation) or DO (Double-floxed Orientation))...recognition sites, its presence induces first opsin inversion, and then lox sites excision, therefore locking... -
Sequencing Options for CRISPR Genotyping
TypeBlog Post...-312. PubMed PMID: 27087594. 16. Tycko, Josh, Vic E. Myer, and Patrick D. Hsu. "Methods for Optimizing...Hendel et al. 2015). Low detection rates mean the investigator can be relatively certain that their samples...off-target mutations are essential data points for investigators looking to definitively link genotype and phenotype...genome-wide unbiased methods. However, most investigators use single cell clones for in vitro CRISPR experiments... -
Mammalian RNAi Tools
TypeCollection...Ventura lab) References Carthew, R. W., & Sontheimer, E. J. (2009). Origins and mechanisms of miRNAs and siRNAs..., M., Rashno, M. M., Abadi, S. J., & Gharagouzlo, E. (2017). Molecular mechanisms and biological functions...Link opens in a new window) . Moore, C. B., Guthrie, E. H., Huang, M. T., Taxman, D. J. (2010). Short hairpin... silencing mechanism. It is a useful tool to investigate the roles of specific genes or to knockdown genes... -
CRISPR 101: Engineering the Plant Genome Using CRISPR/Cas9
TypeBlog Post...:947–951 . https://doi.org/10.1038/nbt.2969 Waltz E (2016) Gene-edited CRISPR mushroom escapes US regulation...homology directed repair, these methods do not involve double strand break (DSB) formation and do not ... -
Recombinase-based State Machines Enable Order-dependent Logic in vivo
TypeBlog Post...implement RSMs? In our study (1), we implement RSMs in E. coli with two plasmids (Figure 3 below): Input Plasmid...then the result of their recombination is the inversion of the DNA between them (Fig. 1A below). In the... -
Antibodies 101: Validation
TypeBlog Post...Bandrowski A, Carr S, Edwards A, Ellenberg J, Lundberg E, Rimm DL, Rodriguez H, Hiltke T, Snyder M, Yamamoto...- it is hard to distinguish between specific, invariable expression and non-specific, background signal... -
Arf GTPase Family
TypeCollection... (Link opens in a new window) Sztul, E., Chen, P., Casanova, J. E., Cherfils, J., Dacks, J. B., Lambright...Santy, L. C., Schürmann, A., Wilhelmi, I., Yohe, M. E., & Kahn, R. A. (2019). ARF GTPases and their GEFs...this list? Fill out our Suggest a Plasmid form or e-mail [email protected] to help us improve this resource...referred to as a minimal triad, with each component involved in GTPase signaling activity. Figure 1: Locations... -
Important Considerations When Using AAVs
TypeBlog Post...Central PMCID: PMC3381869. 7. McCarty, D. M., Paul E. Monahan, and Richard J. Samulski. "Self-complementary...Drugs, DREADDs) and optogenetics as tools to investigate the roles of certain cell types in locomotion...cause the least amount of immune response. Any investigator who is using an in vivo animal model should ... -
Antibodies 101: Introduction to Antibodies
TypeBlog Post...Engineering production of functional scFv antibody in E. coli by co-expressing the molecule chaperone Skp....lot of flexibility in which proteins you can investigate in each panel. An advantage of flow cytometry.... These applications are typically used for investigating signaling pathways or developing therapeutics... -
Tips for a 1st Time CRISPR User (by a 1st Time CRISPR User)
TypeBlog Post...-mediated gene inactivation. Doench JG, Hartenian E, Graham DB, Tothova Z, Hegde M, Smith I, Sullender...growing so slowly, there isn’t much maintenance involved. Also it was really fun to find individual colonies... -
Brzezinski Lab CRISPR Collection
TypeCollection...Goodson, N. B., Park, K. U., Schwanke, M., Office, E., Schneider, S. R., Abraham, J., Hensley, A., Jones...CRISPR plasmids to study the regulatory networks involved in retinal development. Depositor...Brzezinski Lab CRISPR Collection The Brzezinski lab investigates gene regulation in the context of the developing...aide in their studies of the regulatory networks involved in retinal development the lab utilizes a CRISPR... -
COVID-19 Resources
TypeCollection...2 plasmids. The collection contains: Plasmids for E. coli and Yeast expression Spike protein constructs...this list? Fill out our Suggest a Plasmid form or e-mail [email protected] to help us improve this resource...infection, such as ACE2 and TMPRSS2, which are involved in the entry of SARS-CoV and SARS-CoV-2. Find ...protease that primes the SARS-CoV-2 S protein and is involved in virus entry into cells. ( 2 ) FURIN - an enzyme...superfamily, binds to the SARS-CoV-2 S protein and is involved in virus entry into cells. ( 4 ) Cathepsin L -...infectivity.( 6 ) PCP4 - (Purkinje cell protein 4) is involved in beating of airway cilia.( 7 ) Cytokines - SARS-CoV... BSG - Wang, K et al, 2020, BioRxiv SARS-CoV-2 invades host cells via a novel route: CD147-spike protein... -
Fluorescent Protein Guide: Activity Regulation
TypeCollection...this list? Fill out our Suggest a Plasmid form or e-mail [email protected] to help us improve this resource...Plasmids Fluorescent Protein Purpose Principal Investigator Plasmids GFP GFP-dependent transcription factors... -
The CRISPR Software Matchmaker: A New Tool for Choosing the Best CRISPR Software for Your Needs
TypeBlog Post...parameters represent. The best learn-by-doing tool is E-CRISP. The authors have made a huge effort so that...shouldn’t be a surprise that the CRISPR system was involved. Today, that same laboratory enjoys a successful... a clear winner, is there something that we can invest our time into learning and extract the most value... -
CRISPR References and Information
TypeCollection...the Jin-Soo Kim Lab . CRISPR-ERA: CRISPR -mediated e diting, r epression, and a ctivation (Link opens in...repression and activation). Genome support for bacteria ( E. coli, B. subtilis ), yeast ( S. cerevisiae ), worm...John Doench , David Root , and Hyongbum Kim Labs. E-CRISPR (Link opens in a new window) Identifies gRNA...Currently supports over 20 model and non-model invertebrate species. Developed by the Kate O'Connor-Giles... -
Immunology Research Plasmids and Resources
TypeCollection...beta 5 FLJ76359, HLA-DRB HLA-E major histocompatibility complex, class I, E DKFZp686P19218, EA1.2, EA2.1...subfamily 2, group E, member 1 TLL, TLX, XTLL NR2E3 nuclear receptor subfamily 2, group E, member 3 ESCS,...PTGER1 prostaglandin E receptor 1 (subtype EP1), 42kDa EP1 PTGER2 prostaglandin E receptor 2 (subtype ... HLA-JY3, HLC-C, PSORS1 HLA-E major histocompatibility complex, class I, E DKFZp686P19218, EA1.2, EA2.1...MGC9284 CTSB cathepsin B APPS, CPSB CTSE cathepsin E CATE CTSL1 cathepsin L1 CATL, CTSL, FLJ31037, MEP ...cell lectin-like receptor subfamily C, member 3 NKG2-E, NKG2E KLRC4 killer cell lectin-like receptor subfamily...IGHV1-69 immunoglobulin heavy variable 1-69 IGHV1-E, IGHV169, IGHV1E IGHV1-8 immunoglobulin heavy variable... -
Empty Backbones - Choosing Your Perfect Plasmid Backbone
TypeCollection... production of His6-MBP-tagged fusion proteins in E. coli SUMO Protein purification pCIOX - Bacterial ...TEV cleavage pExp-NusA - Expression of protein in E. coli as TEV cleavable N-terminal His and NusA fusions...this list? Fill out our Suggest a Plasmid form or e-mail [email protected] to help us improve this resource...BiFC vector (pEarleyGate201-YN) Zeocin® is an InvivoGen trademark. Return to top Content last reviewed... -
Luciferase Plasmid Collection
TypeCollection...luciferase gene allow for increased copy number in E. coli and expression of luciferase in the cytoplasm...this list? Fill out our Suggest a Plasmid form or e-mail [email protected] to help us improve this resource...212936 pGL3 Basic Vector Firefly Vector for investigating regions controlling transcription Debrya Groskreutz...hRlucCP/Puro) RapidResponse™ Renilla Vector for investigating regions controlling transcription Pete Stecha...212933 pGL4.82(hRluc/Puro) Renilla Vector for investigating regions controlling transcription Pete Stecha...