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We narrowed to 650 results for: CHI;

Showing: 181 - 210 of 650 results

  1. Choosing the B(right)est Fluorescent Protein: Aggregation Tendency

    Type
    Blog Post
    ...thanks to our guest bloggers Joachim Goedhart and Marieke Mastop! Joachim Goedhart is an assistant professor...This post was contributed by guest bloggers Joachim Goedhart and Marieke Mastop from the Section of Molecular...fluorescent probes. You can follow him on twitter: @joachimgoedhart.   Marieke Mastop is a PhD candidate at the...

  2. CRISPR Methods for Bacteria: Genome Engineering, CRISPRa, CRISPRi, Base Editing, and More

    Type
    Blog Post
    ...CRISPR-Cas9-assisted genome editing in Escherichia coli. Acta Biochimica et Biophysica Sinica. https://doi....a technique that utilizes phage recombination machinery to promote homologous recombination of linear ...resistance E. coli carrying the phage recombineering machinery are first electroporated with pCas9. Then, pCRISPR.../10.1038/nbt.3043 Bikard D, Jiang W, Samai P, Hochschild A, Zhang F, Marraffini LA (2013) Programmable...

  3. Hot Plasmids February 2024

    Type
    Blog Post
    ...which uses a much shorter crRNA. Cas12-RNP-PAGE achieved high editing frequency (nearly 100% in some cases...method is fast! These editing frequencies can be achieved with a 30-minute incubation, even with multiplexed...best-performing variant, CAP-Mac, was effective at breaching the blood-brain barrier and delivering a transgene...as well? Our collection includes a mouse and a chimeric rabbit (original mouse Fab region with rabbit ...

  4. Savvy Advocates Needed to Navigate a Scientific Enterprise in Flux

    Type
    Blog Post
    ...Relations Board to allow graduate students who work as teaching or research assistants to join unions. The topic...provide a better deal for students in areas such as teaching salaries, healthcare access, and benefits; however... Genetics Education Project Lisa Coleman, PhD - Chief Diversity Officer Harvard University  The symposium...research: recommendations from the Future of Research Chicago Symposium F1000Research, 2016. Kyle T. Dolan, Joseph...

  5. PRIDICT: Predicting Efficiencies of Prime Editing Guide RNAs

    Type
    Blog Post
    ... efficiency and building prediction tools with machine learning (PRIDICT, PRIDICT2.0).   How to apply...chromatin-based prediction tool (ePRIDICT). In summary, machine learning-based prime editing prediction models ...computational biology. His expertise includes applying machine learning techniques to make sense of biology, with...B. van, Krauthammer, M., & Schwank, G. (2024). Machine learning prediction of prime editing efficiency...

  6. Viral Vectors 101: Voltage Indicators

    Type
    Blog Post
    ...level in vitro. In vivo monitoring has also been achieved with these sensors, but further optimization is...cell level as they are in vitro. Factors like photobleaching, lack of red-shifted indicators, and brightness...electrochromic FRET capable GEVIs, with rhodopsin quenching the added fluor for this two-part fluorescent ...voltage-dependent changes in rhodopsin modulate the degree of quenching of the fluor via FRET. This system is particularly...

  7. Plasmids 101: Degron Tags

    Type
    Blog Post
    ... tag, and the endogenous cellular degradation machinery to function. The dTAG is the smallest tag reviewed... and widespread usage, a proteolysis-targeting chimera (PROTAC) was developed for HaloTag. PROTACs are...functions – there are antibodies to it and ways to achieve fluorescence with it. AID The auxin-inducible degron...441. doi: 10.1038/s41589-018-0021-8 Holland AJ, Fachinetti D, Han JS, Cleveland DW. Inducible, reversible...

  8. Golden Gate Assembly Upgrades: More Fragments, Faster Assembly, and Higher Fidelity

    Type
    Blog Post
    ... more than 10 modules often rely on two-step hierarchical approaches using different Type IIS restriction...Our work demonstrates that it is now possible to achieve 20+ fragment assemblies with both robust efficiency...Additional confirmation of accurate assemblies was achieved by the sequencing of plasmids isolated from blue...fragment lacI/lacZ cassette assembly was easily achievable with high levels of transformants and low backgrounds...

  9. Light Sheet Fluorescence Microscopy

    Type
    Blog Post
    ...enabling microscopes to visualize the cellular architecture of whole tissues in 3D with unprecedented detail... through the specimen, which often leads to photobleaching.  Furthermore, point-by-point scanning along...focus light. Therefore, there is much less photo-bleaching or photo-damage than in conventional laser scanning...1007-1009. Pubmed. H.-U. Dodt, U. Leischner, A. Schierloh, N. Jährling, C.P. Mauch,  K. Deininger, et al...

  10. Antibodies 101: Producing Recombinant Antibodies

    Type
    Blog Post
    ...antibody production is Komagataella phaffii (a.k.a  Pichia pastoris) because it secretes fewer of its own ...include Human Embryonic Kidney (HEK) cells and Chinese Hamster Ovary (CHO) cells. (L'Abbé et al., 2018...-8730-6_1. PMID: 30242676. Frenzel A, Hust M, Schirrmann T. Expression of recombinant antibodies. Front....org/10.1016/j.cocis.2014.10.002. Randolph TW, Schiltz E, Sederstrom D, Steinmann D, Mozziconacci O, Schöneich...

  11. An Introduction to Adenovirus

    Type
    Blog Post
    ... E., Nguyen, J., … Chiu, C. Y. (2023). Adeno-associated virus type 2 in US children with acute severe ...three (Servellita et al., 2023). Normally when children have acute viral hepatitis it is caused by the...noting that it is possible to design a vector that achieves persistent transgene expression in certain cell...

  12. A Career in Grants: How To Become a Grant Professional

    Type
    Blog Post
    ...the term ‘grant writer’, but make no mistake, researching, winning, and effectively managing grants involves...development, certification, degrees, foundations, coaching and even Twitter Chats dedicated to the Grant ..., e.g. education, poverty, environment, early childhood development, or community development? Do you ...consultants who offer grant training, mentoring and coaching services, including Smartegrants, to help those...

  13. Your Lentiviral Plasmid FAQs Answered

    Type
    Blog Post
    ...transfer vectors based on whether or not they have a chimeric 5'LTR. Second-generation transfer plasmids have... while 3rd-generation transfer plasmids have a chimeric 5'LTR that includes a CMV or RSV promoter as well... well as a portion of the 5'LTR. Including a chimeric 5'LTR removes the requirement for the HIV Tat protein...SIN is shorthand for self-inactivating, which is achieved by deleting a large portion of the 3'LTR in the...

  14. Viral Vectors 101: Pseudotyping

    Type
    Blog Post
    ... cell lines that do not express these sugars. (Takeuchi et al., 1997). The alphaGal sugars end up in the...withstand ultracentrifugation which helps scientists achieve a high titer (reviewed in Joglekar and Sandoval... can bind and transduce many cell types before reaching the target cells. VSV-G is also toxic at high ...832 . https://doi.org/10.1182/blood-2001-11-0042 Takeuchi Y, Liong SH, Bieniasz PD, Jäger U, Porter CD, ...

  15. Deciphering the Mysteries of Behavior with Viral Vectors

    Type
    Blog Post
    ...general, or anxiety associated with psychiatric diseases such as schizophrenia or bipolar disorder. In both ...found that downregulating expression of miR124a achieved a similar improvement in the forced swim test....the BDNF hypothesis of depression?." Molecular psychiatry 12.12 (2007): 1079. PubMed PMID: 17700574. Hare...

  16. Plasmids 101: Shuttle Vectors

    Type
    Blog Post
    ... will be recognized by its host’s replication machinery. A shuttle vector designed for unrelated bacteria...requires compatible cell lines that express viral machinery. Alternatively, some slow-growing cells, particularly...nutrient. The vector provides the missing synthesis machinery, and the cells are switched to minimal media, ...transfection into producer cells, which have the machinery to manufacture infectious viral particles. Those...

  17. Plasmids 101: Stringent Regulation of Replication

    Type
    Blog Post
    ...plasmids rely on their host cell's replication machinery in order to replicate—but not always to the same..., as they only require the host's replication machinery for elongation and termination. These types of... folding and blocks RepZ translation (Asano & Mizobuchi, 1998). This system keeps RepZ levels low even...doi.org/10.5772/19313 References Asano, K., & Mizobuchi, K. (1998). Copy number control of IncIalpha plasmid...

  18. Viral Production at Addgene

    Type
    Blog Post
    ... generally two to three fold higher than those achieved using the standard qPCR method.  Titering by the...preparations is assayed by comparing the relative stoichiometric ratios of the viral capsid proteins VP1, VP2...transfer plasmid Addgene uses a rigorous barcode matching system that follows the sample from the DNA tube...    Figure 4: Schematic of the barcode matching system and PCR-based confirmation of transfer ...

  19. CRISPR 101: Targeting RNA with Cas13a (C2c2)

    Type
    Blog Post
    ...genes that fell into 3 categories based on the architecture of the CRISPR protein in question: C2c1, C2c2...single crRNA The Feng Zhang lab used Cas13a from Leptotrichia shahii (LshCas13a) to begin to characterize ...specific RNA sequences from a population (either enriching or depleting specific RNAs out of a pool of RNAs...et al., 2017). They characterized Cas13a from Leptotrichia wadei (LwaCas13a), using a superfolder GFP fusion...

  20. Advanced Uses of Cre-lox and Flp-FRT - A Neuroscientist’s View

    Type
    Blog Post
    ...recombinase have been developed. A common means to achieve ligand control is to fuse a mutant estrogen receptor...be transfected by in utero electroporation. To achieve sparse labeling of individual neurons one could...Central PMCID: PMC41617. 7. Metzger, D, J Clifford, H Chiba, and P Chambon. 1995. “Conditional Site-Specific...Recombination in Mammalian Cells Using a Ligand-Dependent Chimeric Cre Recombinase.” Proc Natl Acad Sci U S A 92(...

  21. Bacterial Toxin-antitoxin Systems as Molecular Biology Tools

    Type
    Blog Post
    ...create unmarked genetic modifications in the yeast Pichia pastoris. By combining both positive and counter...first described by studies of EcoRI and Bsp6I in Escherichia coli in 1995 when introduction of restriction-modification..., Hanna Engelberg-Kulka, and Gad Glaser. "An Escherichia coli chromosomal" addiction module" regulated...counter-selectable marker for unmarked genetic modification of Pichia pastoris." FEMS yeast research 9.4 (2009): 600-...

  22. Antibodies 101: Single Chain Fragment Variables (scFvs)

    Type
    Blog Post
    ... are part of engineered chimeric antigen receptors (CARs). CARs are a chimera of scFvs and T cell receptors...anti-digoxin single-chain Fv analogue produced in Escherichia coli. Proceedings of the National Academy of ...A, Sivaccumar JP, Ruvo M (2020) Evolution of Escherichia coli Expression System in Producing Antibody ...

  23. Antibodies 101: Beyond Surface Labeling

    Type
    Blog Post
    ...1: Indirect detection of target markers can be achieved through two-protein labeling by utilizing(a) biotin-streptavidin...e.g., for cellular activation. In addition, FACS achieves greater purity of your sorted cell sample than...proliferation between cell populations can be visualized, achieved through, for example, a previously induced gene... your marker of interest. Indirect labeling is achieved through protein-tag or protein-protein interactions...

  24. Fluorescent Proteins 101: When GFP lets you down

    Type
    Blog Post
    ...   Many thanks to our guest blogger, Joachim Goedhart! Joachim Goedhart is an assistant professor at ...This post was contributed by guest blogger Joachim Goedart, an assistant professor at the Section of ...fluorescent probes. You can follow him on twitter: @joachimgoedhart.   References 1. Zacharias, David A., et al...

  25. Viral Vectors 101: AAV Serotypes and Tissue Tropism

    Type
    Blog Post
    ...have created many engineered serotypes by mixing, matching, and modifying capsid and genomic structures. ...., Brown, K. E., Walters, R. W., Zabner, J., & Chiorini, J. A. (2001). Adeno-Associated Virus Serotype...Kuleshova, E., Ojala, D., Huang, C.-C., Gerfen, C. R., Schiller, J., Dudman, J. T., Hantman, A. W., Looger, L....M., Keshavjee, S., Brown, K. E., Welsh, M. J., Chiorini, J. A., & Zabner, J. (2001). Binding of adeno-...

  26. Easi-CRISPR: Generating Knock-In and Conditional Mouse Models

    Type
    Blog Post
    ...pseudopregnant female mice. If you obtained the desired chimeric mice, you would breed them many times, hoping ...line with a recombinase. If it turned out your chimeric mice did not contain edited germ cells, your experiment...inject the ES cells into a mouse embryo to create chimeric mice. This laborious process takes ~8-10 months...When delivering the ssDNA template and CRISPR machinery, Quadros et al. recommend ribonucleoprotein (RNP...

  27. Plasmids 101: Repressible Promoters

    Type
    Blog Post
    ...engineered from the cmt and cym bacterial operons. A chimeric transactivator, cTA, was created by fusing CymR...the system, researchers have used split GAL4 architecture or combined the system with Flp/FRT, as reviewed...) to GAL4 or VP16 activation domain results in chimeric GAL80-repressible and GAL80-insensitive activators...lethality in flies. Riabinina et al. also developed chimeric transactivators GALQF and LexAQF, which activate...

  28. Building Global Connections with the International Mentorship Program USA-EUROPE

    Type
    Blog Post
    ...In an era where millions of people hold a PhD, achieving our career goals can be a real challenge. On top... hinder them from advancing their careers and achieving success.” Pedro Gonzalez Menendez (Mentee IMP ...compared to their pre-program self-assesment. Achievements directly attributable to participation in the...interested in breast cancer and metastasis as well as teaching and mentoring activities. Follow her on twitter...

  29. A Practical Guide to Optimizing AAV DIO and FLEx Vector Expression

    Type
    Blog Post
    ...optimal/minimal injection titer and volume needed to achieve adequate expression in your cells of interest.  ...waiting periods and/or higher injection titers to achieve adequate expression. Conducting a time course experiment...to minimize off-target expression while still achieving adequate expression in your cells of interest....different brain region that the same result will be achieved. The fact is that there are many known and unknown...

  30. Fluorescent Proteins 101: Monitoring Cell Mobility Using Fluorescent Proteins

    Type
    Blog Post
    ...2. Fluorescence Activated Cell Sorter (FACs) A machine that sorts cells according to whether or not they... deflect them into appropriate containers. The machine is useful for distinguishing cell populations that...21596592. PubMed Central PMCID: PMC3164316. 5. Chieppa, Marcello, et al. "Dynamic imaging of dendritic...165. PubMed PMID: 18193049.  9. Filipe-Santos, Orchidée, et al. "A dynamic map of antigen recognition ...
Showing: 181 - 210 of 650 results