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We narrowed to 185 results for: INV-E

Showing: 41 - 80 of 185 results

  1. Antibodies 101: Conjugation

    Type
    Blog Post
    ...doi.org/10.1016/j.it.2012.02.010 Dennler P, Fischer E, Schibli R (2015) Antibody Conjugates: From Heterogeneous...doi.org/10.3390/antib4030197  Hegazy M, Cohen-Barak E, Koetsier JL, et al (2020) Proximity Ligation Assay...Reisfeld RA (1988) Determination of the Number of e-Amino Groups Available for Conjugation of Effector...However, there are certainly other approaches that involve modifying the antibody sequence or that rely on...

  2. The AAVantages of AAV in CRISPR Screens

    Type
    Blog Post
    ...Volak, A., Spirig, S. E., Muller, A., Sousa, A. A., Tsai, S. Q., Bengtsson, N. E., Lööv, C., Ingelsson...10.1101/2023.06.13.544831 Santinha, A. J., Klingler, E., Kuhn, M., Farouni, R., Lagler, S., Kalamakis, G....less work for the first two steps, as no mice are involved. Ex vivo starts with transducing cells with the...lentivirus requires a BSL-2 setup. This requires a less involved setup when working with mice and slightly less...high degree. This is particularly helpful when investigating tissue-specific diseases, as you can target ...like Cas12a (Cpf1). Even more creative options involve generating strains of mice that stably express ...

  3. SARS-CoV-2/COVID-19 Detection Methods Based on CRISPR/Cas

    Type
    Blog Post
    ...isothermal amplification by primers that target the E and the N2 gene of SARS-CoV-2. If the Cas12a-sgRNA...CRISPR-based SHERLOCK (Specific ​H​igh ​s​ensitivity ​E​nzymatic ​R​eporter un​LOCK​ing) technique Sensitivity...Granados A, Sotomayor-Gonzalez A, Zorn K, Gopez A, Hsu E, Gu W, Miller S, Pan C-Y, Guevara H, Wadford DA, Chen...doi.org/10.1101/2020.02.29.971127 Rauch, J. N., Valois, E., Solley, S. C., Braig, F., Lach, R. S., Baxter, N...University. Shravanti Suresh is a PhD candidate who investigates cellular aspects of CRISPR/Cas immune systems...

  4. Mobile-CRISPRi: Bringing CRISPRi to Diverse Bacteria

    Type
    Blog Post
    ...thetaiotaomicron, Mycobacterium, Pseudomonas, and E. coli. However, many non-model bacterial species lack...100 to 1,000 colonies, but some bacteria, including E. faecalis and P. mirabilis, had only ~1 transfer per... PubMed Central PMCID: PMC6424567. Peters, Joseph E. "Tn7." Mobile DNA III. American Society of Microbiology...into multiple bacterial species including several involved in human disease. However, the rate of transfer...for the study of essential genes such as those involved in antibiotic resistance. Are you ready to move...

  5. Lentiviral CRISPR Libraries Enable Genome-Scale, Knockout Screening

    Type
    Blog Post
    ...Screening in Human Cells. Shalem O, Sanjana NE, Hartenian E, Shi X, Scott DA, Mikkelson T, Heckl D, Ebert BL, ...CRISPR-Guide RNA Library. Koike-Yusa H, Li Y, Tan E-P, Del Castillo Velasco-Herrera M, Yusa K. Nature ...melanoma model, they also went in search of genes involved in resistance to the cancer drug vemurafenib. ...lab has used their library to screen for genes involved in bacterial toxin resistance, identifying 4 previously...

  6. Using AAV for Neuronal Tracing

    Type
    Blog Post
    ...PubMed Central PMCID: PMC2600441. Foust, K.D., Nurre, E., Montgomery, C.L., Hernandez, A., Chan, C.M. and ...19098898. PubMed Central PMCID: PMC2895694. Hollis, E. R. 2nd, Kadoya, K., Hirsch, M., Samulski, R. J., ...S.F., Andrews, A.M., Lutton, E.M., Mu, D., Hudry, E., Hyman, B.T., Maguire, C.A. and Ramirez S.H. (2017... Wakeman, W., Li, Y., Feng, D., Ho, A., Nicholas, E., Hirokawa, K.E., Bohn, P., Joines, K.M., Peng, H....Central PMCID: PMC5102064. Roberts, T. F., Hisey, E., Tanaka, M., Kearney, M., Chattree, G., Yang, C. ... Ritola, K.D., Lindo, S., Michael, S., Kuleshova, E., Ojala, D., Huang, C.C., Gerfen, C.R., Schiller, ...hippocampus and the prefrontal cortex brain regions is involved in the formation of episodic memory, a special...

  7. Plasmids 101: Shuttle Vectors

    Type
    Blog Post
    ...Plant vectors are usually designed to shuttle between E. coli and Agrobacterium tumefaciens, a plant-infectious...across species, for example, in both S. cerevisiae and E. coli. Auxotrophic selection is especially common ... shuttling between. For example, if you are using E. coli simply for easy cloning and quick DNA production...vectors? The production of viral vectors typically involves shuttle vectors, also sometimes called transfer...target cells. However, while shuttle vectors are involved in the production of viral vectors, we consider...

  8. What's the Best Way to Elute and Store Your Plasmid DNA?

    Type
    Blog Post
    ...during plasmid DNA extraction and purification from E. coli is the presence of the protein endonuclease ...encoded by the gene endA. Nowadays, the most common E. coli strains used for plasmid amplification are endA...DNA preparation in any downstream application involving enzymatic reactions, a valid alternative can be...

  9. Illuminating Choices: A Guide to Selecting Fluorescent Dyes and Ligands

    Type
    Blog Post
    ..., R., Zuber, J., Connelly, S. M., Mathew, E., & Dumont, M. E. (2014). Fluorescent approaches for understanding...Jeddi-Tehrani, M., Mahmoudi, A. R., Bayat, A. A., Shaban, E., Vafakhah, M., Darzi, M., Tarahomi, M., & Ghods, ...Bhattacharya, K., Pyrgiotakis, G., Zhang, Z., Purington, E., Gramlich, W., Grondin, Y., Rogers, R., Bousfield...Stability of fluorescence: For applications that involve ranges of pH, and significant lengths of time, ...incorporation of fluorophores, as seen in studies involving dopamine D2/D3 receptors (Allikalt et al., 2020...processes like mitochondrial dynamics. This process, involving mitochondrial homeostasis maintained through mechanisms...

  10. History of CRISPR Cas - A tale of survival and evolution

    Type
    Blog Post
    ...the CRISPR sequences were initially discovered in E. coli in 1987 (1), the concept that these clustered... K, Haft DH, Barrangou R, Brouns SJJ, Charpentier E, Horvath P, Moineu S, Mojica FJM, Wolf YI, Yakunin...various cas genes not only to store a record of invading phages but to also destroy the phage upon re-exposure... the foreign DNA, a cut is made to destroy the invader. Wikipedia, accessed 25 November 2013. Author...systems provides powerful lines of defense against invading phages. Rising to the challenge - Evolution of...research. Do you have a cool idea or application that involves CRISPR/Cas technology? Let us know.  References...

  11. A Look Back at One Year of Plasmid Sharing for COVID-19 Research

    Type
    Blog Post
    ...Biering SB,  Sarnik SA, Wang E, Zengel JR, Sathyan V, Nguyenla X, Van Dis E, Catamura C, Yamashiro LH,  ...Puschnik AS, Bertozzi CR, Carette JE, Stanley SA, Harris E, Konermann S, Hsu PD (2021) Genome-wide, bidirectional...starting point for SARS-CoV-2 researchers to investigate how the virus interacts with human cells. Addgene...

  12. Tips from the Repository Trenches: Using Barcodes to Track Samples

    Type
    Blog Post
    ...different strains of E. coli that can be used for transformation. The preferred E. coli strains for storage... DNA has been received and stored, it is put into E. coli strains that have been treated to easily take...kanamycin resistance gene. Every step along the way involves good aseptic technique and controls to ensure ...

  13. Special Delivery: Fluorophore Targeting for FRET Studies

    Type
    Blog Post
    ...amino acids E. coli and Mammalian Cells pEVOL-pAzF synthetase for p-azido-l-phenylalanine E. coli pEVOL-pBpF...has proven most effective only in a few cell types (E. coli and Xenopus oocytes, mainly). Nevertheless, ...pEVOL-pBpF synthetase for p-benzoyl-l-phenylalanine E. coli pAcBac1.tR4-MbPyl pyrrolysyl-tRNA synthetase...commercially available as Lumio Green and Lumio Red from Invitrogen (now ThermoFisher Scientific). These reagents...structurally characterizing large ion channels involved in skeletal muscle contraction.   References 1...

  14. Plasmids 101: Dimers and Multimers

    Type
    Blog Post
    ...plasmid multimerization in the naturally occurring E. coli plasmid ColE1 (Goebel & Helinski, 1968; Bazaral... (Bedbrook & Ausubel, 1976; Fishel et al., 1981). E. coli strains such as DH5α, HB101, NEB Stable, Stbl3...multiple circular DNA forms of colicinogenic factor E-1 from Proteus mirabilis. Biochemistry. 1968 Oct;7...1566524020666200427110452. PMID: 32338220. Crozat E, Fournes F, Cornet F, Hallet B, Rousseau P. Resolution...during our quality control process. Since MiSeq involves sequence assembly from short reads (<300 bp) which...

  15. New and Upcoming Viral Vectors - May 2020

    Type
    Blog Post
    ...Lentiviral preps for expressing the E protein (pLVX-EF1alpha-SARS-CoV-2-E-2xStrep-IRES-Puro) and NSP2 (pLVX-EF1alpha-SARS-CoV...service in 2016, we’ve added many new tools to our inventory of ready-to-use viral vectors. Here are some of...interneurons (83895). See our Serotype Testing AAV inventory for the entire collection! Plasmid Serotype...Fishell   Chemogenetics Chemogenetics offers noninvasive methods for manipulating cell activity, particularly...

  16. Top Requested Lentivirus and AAV of 2016

    Type
    Blog Post
    ...Ye C, Rogan SC, Adams AC, Cusher DS, Maratos-Flier E, Roth BL, Lowell BB. Rapid, reversible activation ...PubMed Central PMCID: PMC3069789. 3. Kügler S, Kilic E, Bähr M. Human synapsin 1 gene promoter confers highly...few inventory items offered domestically, but by the end of 2016, we expanded our viral inventory to 25... lentiCas9-Blast Most of Addgene’s lentiviral inventory is based on CRISPR tools, though we do provide...pAAV-hSyn-DIO-hM4D(Gi)-mCherry Our current AAV inventory consists almost entirely of viruses expressing... Carolina, Chapel Hill. These can be used to investigate the effects of activating particular signaling...neurons drives feeding behavior in mice. J Clin Invest. 2011 Apr;121(4):1424-8. PubMed PMID: 21364278....

  17. Proximity Labeling: A Powerful Tool for Protein Complex Purification and Proteomic Mapping

    Type
    Blog Post
    ...Burke Lab (Roux et al., 2012). BioID utilizes the E. coli biotin ligase, BirA*, with a site mutation (...DNA-binding domain (233 amino acids) found in the E. coli version. It also contains a single mutation ...signal to noise ratio 30-fold compared to the standard E. coli BirA* mutant. RapID consists of this mutant ...Interactions through Spatial Proximity." Journal of Investigative Dermatology 137.12 (2017): e197-e203. PubMed...

  18. Plasmids 101: Expressing shRNAs for RNA Interference

    Type
    Blog Post
    ...00045-5 De Rie, D., Abugessaisa, I., Alam, T., Arner, E., Arner, P., Ashoor, H., Åström, G., Babina, M., Bertin... S., Montgomery, M. K., Kostas, S. A., Driver, S. E., & Mello, C. C. (1998). Potent and specific genetic...15476286.2023.2288741 Taxman, D. J., Moore, C. B., Guthrie, E. H., & Huang, M. T. (2010). Short hairpin RNA (SHRNA...silencing. RNA interference RNAi is a useful tool to investigate the roles of specific genes or to knock down ...term for a number of different applications that involve different types of small RNA molecules, such as...

  19. Plasmids 101: Modular Cloning Applications and Kits

    Type
    Blog Post
    ... a supplemental add-on kit and is compatible with E. coli and/or cell-free systems. Protein expression... et al. “EcoFlex: A Multifunctional MoClo Kit for E. Coli Synthetic Biology.” ACS Synthetic Biology, vol...CIDAR MoClo: Improved MoClo Assembly Standard and New E. Coli Part Library Enable Rapid Combinatorial Design...//doi.org/10.1021/acssynbio.6b00337. Lee, Michael E., et al. “A Highly Characterized Yeast Toolkit for...model organisms and backgrounds, MoClo can be invaluable for more complex editing and screening experiments...

  20. Targeted Mutagenesis with EvolvR

    Type
    Blog Post
    ...polymerase I (PolI3M) is capable of mutagenesis in E. Coli, but this approach requires special bacterial...mutations per nucleotide per generation of wild-type E. coli, while only increasing the standard mutation...be used to map protein-protein interactions, investigate non-coding segments of the genome, or engineer...

  21. PiggyBac-ing Through the Genome Editing Field

    Type
    Blog Post
    ...and strategies for their projects. References 1. L. E. Woodard and M. H. Wilson, “piggyBac-ing models and...Biol., Apr. 2016. PubMed PMID: 27075976. 4.  X. Li, E. R. Burnight, A. L. Cooney, N. Malani, T. Brady, J.... Menon, G. S. Parker, S. J. Qualls, B. M. Lewis, E. Ke, C. T. Dargitz, R. Wright, A. Khanna, F. H. Gage... likely due to the fact that PBx relies on the inverted terminal repeats (ITRs) found on the piggyBac™... Grégoire and M. Kmita, “Recombination between inverted loxP sites is cytotoxic for proliferating cells...

  22. CRISPR in the Clinic

    Type
    Blog Post
    ...infections. The Cas3 enzymes target several strains of E. coli and lethally chop their genomes into many, many...Hussein M, Molina MA, Berkhout B, Herrera-Carrillo E. A CRISPR-Cas Cure for HIV/AIDS. Int J Mol Sci. 2023...strategies for CRISPR therapies. Systemic therapies involve CRISPR-Cas delivery directly to the patient. Patient-derived...returned to the patient. Finally, allogenic therapies involve editing healthy donor-derived cells and delivering... treatment strategies for these disorders have involved turning on fetal hemoglobin to compensate for ...returning them to the patient. The first human trial involving CRISPR Cas9 for these diseases was sponsored by...evolving, and many of the challenges with therapies involve patient safety, effectiveness, and delivery. Patient...

  23. Transgenic Organisms, Cas9 Gene Drives, and Appropriate Safeguards

    Type
    Blog Post
    ...345, 626–628 (2014). PubMed. Gantz, V. M. & Bier, E. The mutagenic chain reaction: A method for converting... doi:10.1126/science.aaa5945. PubMed. DiCarlo, J. E., Chavez, A., Dietz, S. L., Esvelt, K. M. & Church...damage public trust in scientists, experiments involving potential gene drives should be conducted with...strategies are available? Molecular confinement involves building gene drives that can spread through populations... been tested in yeast5. Ecological confinement involves performing experiments in a geographic area where...relevant mosquito species. Reproductive confinement involves working with laboratory organisms that can't reproduce...publications. Thank you to our guest blogger! An inventor of technologies that harness evolution, Kevin ...

  24. GCE4All: Making Genetic Code Expansion Accessible

    Type
    Blog Post
    ... In the late nineties, this was developed into an E. coli system, with the noncanonical amino acid in ...expression of phosphoserine-including proteins in E. coli (Zhu et al., 2022). It uses the TAG codon for...components, meaning that the final protocol may involve a sensitive balance of trade-offs. This balance...

  25. Genome-wide Screening Using CRISPR

    Type
    Blog Post
    ...carries CRISPR libraries that target mouse, human, fly, E. coli, and T. gondii genes.  2) What genetic modification... are identified using next-generation sequencing (E). Considerations and tips for successful screens... lead to a wealth of additional experiments investigating the role of individual genes or entire pathways...a population of mutant cells to identify genes involved in a particular phenotype. CRISPR can be readily...specific classes of human genes. What are the steps involved in a CRISPR screen? Performing a forward genetic...

  26. The 12 Days of CRISPR

    Type
    Blog Post
    ...On the 11th day of CRISPR, Addgene gave to me: Cas-E-le-ven! (Cas11)🎵 On the 10th day of CRISPR, Addgene... the 12th day of CRISPR, Addgene gave to me: Noninvasive assessment of gut function using transcriptional...

  27. Addgene: The First Twenty Years

    Type
    Blog Post
    ...request from — Addgene!  Addgene was established as an e-commerce company, which helped it stay ahead of the...for now-common business practices, like accepting e-signatures and designing a website users could easily...hear it all the time, but Addgene is a totally invaluable resource. I really appreciate the work you do...

  28. INTEGRATE: Bacterial Genome Engineering Using CRISPR-Transposons

    Type
    Blog Post
    ... were used to express the necessary components in E. coli (Klompe et al., 2019). Recently, the lab introduced...temperatures. Data adapted from Vo et al., 2020. In E. coli, the new streamlined VchINT system was shown...programmability: depending on the transposases involved, they insert either randomly, or at fixed genomic... L ends).  Figure 1: (A) Components involved in each type of CRISPR-Transposon system. (B) ... a streamlined expression approach (Fig. 2A), involving two-plasmid or single-plasmid systems (Vo et al...insertions are not currently possible, unlike methods involving homologous recombination. Additionally, these ...

  29. CRISPR Library Amplification

    Type
    Protocol
    ...Maxipreps rely on incremental, ordered cell lysis. E. coli cells are subject to lysis by freeze-thaw if...pre-chilled cuvette. Pulse. Caution Electroporation involves the use of high voltages, please use caution when...

  30. AAV Vector Quality Control: Going the Extra Mile with NGS

    Type
    Blog Post
    ...Bourges D, Ersing I, Haery L, Harten DeMaio K, Sanders E, Tasissa M, Kostman M, Tillgren M, Makana Hanley L...678 . https://doi.org/10.1089/hum.2019.277 Lecomte E, Tournaire B, Cogné B, Dupont J-B, Lindenbaum P, Martin-Fontaine...and it may also be aligned to plasmids in our inventory containing the suspected contaminating sequence...

  31. Synthetic Biology, Artificial Chromosomes, iGEM & More - Interview with Tom Ellis

    Type
    Blog Post
    ...projects underway in the Ellis lab including yeast and E. coli engineering, microbial cooperation, and antibiotic...team also investigates bacteria and extremophiles for biotechnology and has been directly involved in supervising... through Open Access. Chapter 7/Outro: Tom’s Involvement with iGEM and Addgene  Many thanks to our guest... He returned to academic research in 2005 to investigate synthetic biology at Boston University, working...

  32. New Optimized Genome-wide CRISPRko, CRISPRi, and CRISPRa Libraries

    Type
    Blog Post
    ...screens." bioRxiv (2018): 356626. 6. Tanenbaum, Marvin E., et al. "A protein-tagging system for signal amplification...Resources Read about CRISPRi and CRISPRa from the NIH E-newsletter ...-wide screens to effectively and efficiently investigate gene function. CRISPR libraries can be used to...

  33. How to Deposit Your Plasmids with Addgene

    Type
    Blog Post
    ...us your plasmid information in spreadsheet format, e-mail us at  [email protected] Special cases Please...can distribute your bacterial strains if they: are E. coli that can be used under BSL1 conditions were...allow you to produce large amounts of your insert in E. coli, then this is a Bacterial Expression vector....deposit, associate the plasmids with a Principal Investigator (PI), provide us with your shipping address ...

  34. Podcast: Interview with Addgene's Founders

    Type
    Blog Post
    ... UBMTA Chapter 4 - Developing Addgene as a modern e-commerce company and establishing credibility Chapter...Operating Officer. Prior to Addgene, Kenneth was an investment banking analyst at Credit Suisse’s Technology...

  35. Performing In Vivo CRISPR Screens Using the FITS Approach

    Type
    Blog Post
    ...to cancer.   References and resources Milling, L. E., Markson, S. C., Tjokrosurjo, Q., Derosia, N. M.,...pRDA_526 vector to knock out 30 genes putatively involved in CD8+ T cell responses to cancer. The screen...

  36. Evolution of Brainbow: Using Cre-lox for Multicolor Labeling of Neurons

    Type
    Blog Post
    ...Matho KS, Ieng S, Fouquet S, Dupin E, Benosman R, Chédotal A, Beaurepaire E, Morin X, Livet J. Neuron. 2014...2.1 cleverly combines Cre-mediated deletion and inversion. The Brainbow-2.1 construct can express one of... or M-CFP.) The construct contains two tandem invertible segments, each with two fluorescent protein coding...color possibilities. The remaining cassette can invert as long as Cre remains expressed; once Cre is removed...recombination is required to prevent constant inversion of the construct. Sanes and Lichtman used pronuclear...

  37. Viral Vectors 101: Integrase-Deficient Lentiviral Vectors

    Type
    Blog Post
    ...Thornhill, S. I., Parsley, K. L., Staal, F. J., Gale, R. E., Linch, D. C., Bayford, J., Brown, L., Quaye, M.,...Leavitt, A. D., Robles, G., Alesandro, N., & Varmus, H. E. (1996). Human immunodeficiency virus type 1 integrase... IDLVs were first discovered by researchers investigating HIV integrase mutants in the early 1990s (Leavitt...wish you the best of luck with any experiments involving IDLVs. And remember: You can never trust a lentivirus...of SCID-X1 patients. The Journal of Clinical Investigation, 118(9), 3143–3150. https://doi.org/10.1172/...

  38. Twenty Years of Sharing: Addgene's Viral Vector Service

    Type
    Blog Post
    ....1007/978-1-0716-2918-5_5 Haery, L., Deverman, B. E., Matho, K. S., Cetin, A., Woodard, K., Cepko, C.,....2019.00093 McDade, J. R., Waxmonsky, N. C., Swanson, L. E., & Fan, M. (2016). Practical Considerations for Using...This process builds on our former approach and involves reading primary literature, soliciting information...

  39. Antibodies 101: Reading a Flow Plot

    Type
    Blog Post
    ...Anfinan, N., Al-Wasiyah,  mohammad khalid, Huwait, E., Gari, M., & Al-Qahtani, M. (2018). Evaluation of..., Ma, M., Sahay, G., Chiu, A., Bader, A., Langan, E., Siebert, S., Li, J., Greiner, D., & Anderson, D....sense once you realize events are plotted on an invisible Cartesian plane (right.) Each event is plotted...

  40. Fluorescent Proteins 101: History of Fluorescent Proteins

    Type
    Blog Post
    ...Martin Chalfie expressed GFP in exogenous organisms (E. coli, and later C. elegans) for the first time. After...ring that glows with a faint green light. To investigate this green glimmer, Shimomura collected many,... in learning more about fluorescent proteins, I invite you to download our Fluorescent Protein eBook and...
Showing: 41 - 80 of 185 results