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We narrowed to 792 results for: LAT

Showing: 151 - 180 of 792 results

  1. All in a Twist: dsRNA

    Type
    Blog Post
    Published
    Feb. 2, 2023, 2:15 p.m.
    ...Transcription, translation, and prone to degradation – those are the words that describe RNA! Double ...transcription. In an uninfected cell, dsRNA rarely circulates, but upon viral infection, this nucleic acid ... RNAi is designed against), RISC can repress translation and/or degrade target mRNA. These RNA tools ... almost always epigenetically silenced, but dysregulation of these systems can lead to local dsRNA production...source of dsRNA (that doesn’t involve cellular dysregulation) is the mitochondria. These organelles have ...During transcription, RNA is synthesized from a DNA template, inherently generating RNA:DNA intermediates in...which suppress endogenous dsRNA formation are dysregulated, immune responses can mount in the absence of...

  2. A History of Genome Engineering in Popular Culture

    Type
    Blog Post
    Published
    Feb. 25, 2020, 2:15 p.m.
    ...revolution in genetic engineering techniques is a speculation of yesteryear which has been realized recently...explained how the four letters of the DNA get translated into the twenty amino acids. Once scientists ...DNA encoded all the information necessary for regulating and propagating life, there was no turning back... molecular biologist was to devise methods to manipulate it. The term genetic engineering itself is attributed...what would happen if these corporations would extrapolate genetic engineering to exploit human beings. ...repaired by homologous recombination with a repair template.  Each zinc finger recognizes a nucleotide triplet...advertisement of the technology on various non-scientific platforms, CRISPR has also transcended into the world of...

  3. Common Injection Routes in Mice

    Type
    Blog Post
    Published
    Jan. 22, 2025, 9:42 p.m.
    ... as “instillation” or “gavage,” as they do not require any punctures. Intratracheal instillation Intratracheal...Skill required: advanced Intranasal instillation Intranasal instillation is very similar to its intratracheal...are used for a broad range of substances, as it circulate what is injected throughout the body. Tail vein...not recommended for tumorigenic compounds, as circulation is slower, and more institutional oversight is...the muscles. The most common injection sites are flat and easily accessible, like on the back. Depending...performed anywhere on the body, somewhere large, flat, and easily accessible (like the back) is preferred...Intratracheal instillation is used to deliver substances directly to the lungs. It is an advanced technique...

  4. Chromoproteins: Colorful Proteins For Molecular Biology Experiments

    Type
    Blog Post
    Published
    Feb. 4, 2020, 2:15 p.m.
    .... Eight years later, scientists isolated pink, purple, and blue chromoproteins isolated from reef-building...reach zooxanthellae, the coral’s symbiotic dinoflagellates that are photosynthetic (Salih et al., 2000...experiment worked just by looking at the cells on the plate. For example, if your plasmid contains a gene encoding...directly use in bacteria, where most genetic manipulations start. Anthony Forster’s lab used codon optimization... Chromoprotein expression from bacteria on agar plate. Image: Liljeruhm et al., 2018. For each chromoprotein...exchange. These vectors contain a toxin gene (regulated by a rhamnose-inducible promoter) and a chromoprotein...Dove SG, Takabayashi M, Hoegh-Guldberg O (1995) Isolation and Partial Characterization of the Pink and Blue...

  5. Light Sheet Fluorescence Microscopy

    Type
    Blog Post
    Published
    July 16, 2015, 2:30 p.m.
    ...solution of benzyl alcohol and methyl salicylate which was later modified by others to produce Murray’...major advances in light microscopy, particularly related to the neurosciences.  These developments in microscopy...single gold particles with diameters less than 4nm. Lateral view of a cleared rat pons, medulla and the first...using optical projection tomography (OPT)  (12) and later Hans-Ulrich Dodt’s lab pioneered the use of light...) (6).   The Dodt and Frank Bradke laboratories later developed the 3DISCO (3-dimensional imaging of solvent...tetrahydrofuran-based tissue clearing (13,14). A few years later, Karl Deisseroth’s lab developed CLARITY, which ...relied on the use of transgenic mice in which a subpopulation of neurons are brightly labeled with GFP (e....

  6. Tips for arabidopsis transformation

    Type
    Blog Post
    Published
    Oct. 25, 2018, 1:23 p.m.
    ... and whether you are inoculating from a frozen stock or a colony from an LB plate. In my hands, a 5 mL...mL starter culture inoculated from a colony on a plate will reach an optimal density in 18-24 hours. I...Arabidopsis, from studying transcriptional and translational dynamics of genes and proteins in living plants... seeds from the pool you will be selecting from later. The Arabidopsis transformation process The process... T1 plants from each transformation. I like to isolate a minimum of five. You have no control over how..., check out Harrison et al., 2006. Tips: Don’t plate seeds too densely during selection: this will increase...difficult. Don’t leave your plants on selective plates for too long: even resistant plants will be affected...

  7. Uncovering Ancestral Bias in CRISPR Research

    Type
    Blog Post
    Published
    Nov. 29, 2022, 2:15 p.m.
    ...tumors, which can lead to new cancer targets and related biomarkers . Data from such screens are often uploaded...Unexpected results Their results showed 96 strongly correlated genetic dependencies that were clearly associated... they looked for a biological explanation, a relationship between the genetic dependencies they identified... ancestries. While there were some exciting relationships identified, 48% were found to be artificial,...bias into their experiments instead of a true correlation.   It turned out there was indeed a bias, and...diversity of genetic variation across the human population. Rather, it primarily reflects European ancestry... to guides in the Broad’s Genomic Perturbation Platform Avana library (used by the Cancer Dependency Map...

  8. CRISPR Antimicrobials

    Type
    Blog Post
    Published
    May 3, 2016, 2:30 p.m.
    ...their resistance genes throughout the bacterial population. But what if we could specifically target only...successfully resensitized a beta-lactam resistant population to the antibiotic, despite this resistance gene...plasmid-based sequences (red). Treating a mixed population, represented here by cells of different colors...given genomic or plasmid sequence from a mixed population without affecting the other bacteria. Targeting...small pools of antibiotic resistant cells could repopulate the environment after treatment. The phagemid...antibiotics can be used to target the sensitive population. Yosef et al.’s proof-of-concept work targeted...infected by the lysogenic phage, rendering the population sensitive to antibiotics. Created with BioRender.com...

  9. Interview: Hodaka Fujii on enChIP, New CRISPR Tools, and More

    Type
    Blog Post
    Published
    Dec. 2, 2014, 7:23 p.m.
    ...transcription and epigenetic regulation. To elucidate molecular mechanisms of regulation of genome functions, ...mechanisms of genome functions such as epigenetic regulation and transcription by using locus-specific chromatin...manner. To achieve this goal, it is necessary to isolate specific genomic regions while retaining molecular... step of knocking-in. In 2013, we showed that isolation of a single copy genomic locus is feasible by ...enChIP, specific genomic regions can be easily isolated for identification of associated proteins, RNAs...that enChIP will markedly accelerate research of regulation mechanisms of genome functions in the hope of... of the reasons why we have only three biology-related Nobel prizes, out of which only one was done in...

  10. Sonic Hedgehog? Sleeping Beauty? Learn About the Genes Behind Addgene’s Conference Room Names

    Type
    Blog Post
    Published
    April 2, 2019, 1:44 p.m.
    ...hedgehog signalling pathway, which is a key regulator of all bilateral animal development. SHH plays a key role...Cancer Institute! Sonic Hedgehog, an important regulator of embryonic development The largest conference...publication, Ivics et al 1997. Spaghetti Squash, a regulator of cellular contraction You may think that a gene...component of the fly actomyosin cytoskeleton that regulates cellular contraction. This network is composed...vessels). Spaghetti squash (Sqh) is the Drosophila Regulatory Light Chain (RMLC) of nonmuscle myosin II (Karess... Jordan and Karess, 1997), which contains the regulatory regions that are necessary for myosin activity...Roger Karess. "Myosin light chain–activating phosphorylation sites are required for oogenesis in Drosophila...

  11. Plasmids 101: Shuttle Vectors

    Type
    Blog Post
    Published
    May 20, 2025, 1:15 p.m.
    ...has seen before, showing how DNA sequences are translated into amino acids. And I was astonished that the...basis of shuttle vectors, which can function in unrelated host species, even from completely different kingdoms...eukaryotic cells, but would prefer to grow, clone, and manipulate their vectors in fast-growing bacteria first....consider how to keep it there. In fast-growing populations of cells, the vector needs to replicate alongside...alongside its host, or it will be lost from the population. A bacterial chromosome has a single origin of...replication machinery. A shuttle vector designed for unrelated bacteria should include oris for both target species...selection, but these resistance systems don’t always translate to eukaryotes. For example, beta-lactam antibiotics...

  12. Antibodies 101: Immunoprecipitation

    Type
    Blog Post
    Published
    Dec. 7, 2021, 2:15 p.m.
    ... protein is up or downregulated, examine a protein’s stability or post-translational modifications, or...aureus. Its isolation by affinity chromatography and its use as an immunosorbent for isolation of immunoglobulins...immobilized antibodies, or immunoglobulins, to isolate a specific protein out of a complex mix. Using ...suitable for all applications, such as protein isolation from serum. In this case, Protein A and Protein... assay, an antibody against the tag is used to isolate the protein of interest. The wide availability ...experiments using a tagged protein may not be translatable to the endogenous system and will need to be...modification. Similarly, RNA IP (RIP) can be used to isolate proteins that bind to RNA, though the details of...

  13. Four Base Editing Reporters to Monitor and Enrich Editing in Real-time

    Type
    Blog Post
    Published
    July 7, 2020, 1:15 p.m.
    ...identification system for the isolation of base-edited cell populations. To develop this method the lab... affecting protein translation of different reporter proteins. Protein translation requires the start ...edited cells without the need for single cell isolation and downstream sequencing (Standage-Beier et al...and Brafman lab showed that GFP positive cells isolated with TREE have a significantly higher frequency...base-editor (C>T conversion) they could induce the translation of any detectable protein, not just fluorescent...base-editing reporter. As the initiation of protein translation at the start codon ATG is universal, the Dow ...Dow lab successfully used GO to induce the translation of an array of different reporters including mScarlet-I...

  14. Harnessing Bacterial Toxins for Allelic Exchange

    Type
    Blog Post
    Published
    Aug. 15, 2019, 12:30 p.m.
    ...recipient, the only antibiotic resistant colonies isolated are those where the allelic exchange vector has...that contained one of eight bacterial toxin genes isolated from E. coli or Psuedomonas aeruginosa. These ...the transcriptional activator rhaS. To allow manipulation of bacteria without an endogenous rhamnose regulon...synthetic ribosome binding site (using the online calculator from the Salis group) (Espah Borujeni et al.,...designed with the help of the DNA tuner program) (Latynski and Valentovich 2014). We routinely clone the ...facilitate the cloning of toxic products. To allow manipulation of naturally chloramphenicol-resistant strains...Anirudh S. Channarasappa, and Howard M. Salis. "Translation rate is controlled by coupled trade-offs between...

  15. Antibodies 101: Normalization and Loading Controls for Western Blots

    Type
    Blog Post
    Published
    April 8, 2025, 1:15 p.m.
    ...intensity. Researchers use later steps in this chain (blue) to infer or calculate things about earlier ones... normalization and loading controls needed for relative quantification of a western blot — and why you...can you accurately compare different lanes? In relative quantification, the POI bands are normalized to...protein concentration in your sample would directly correlate to protein abundance on your membrane, which would...multiple sources of error (red) influence these relationships and must be addressed with separate controls...accurately quantified In western blotting, the relationship between protein abundance and band intensity... BioRender.com. To see why this S-shaped relationship poses problems for proteins of very different...

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

    Type
    Blog Post
    Published
    June 3, 2021, 1:15 p.m.
    ...phenotype. Incubating the phage display library with plate-bound antigen helps select for high affinity scFvs...bacteriophages. This library is incubating with plate-bound antigen to select for scFvs with high affinity...the non-oxidizing cytoplasm of bacteria, scFvs accumulate in insoluble inclusion bodies. While it’s possible... has led to their application for basic and translational research.  Basic research applications of scFvs...generate since it’s genetically encoded.   Translational applications of scFvs: Just like antibodies...scFvs are also used for eye diseases, such as age-related macular degeneration (AMD) since direction injection...cell. When a CAR has an scFv that binds cancer-related antigens, binding of the scFv part of the receptor...

  17. CRISPR Challenges: Standardization and Homology Directed Repair

    Type
    Blog Post
    Published
    Feb. 14, 2018, 2:36 p.m.
    ...with ssDNA repair templates. Their work demonstrates that different repair template strategies activate...using dsDNA templates achieving ~1-10% efficiency. Easi-CRISPR uses a long ssDNA template to increase ...These complementary goals are aimed at improving translation of CRISPR technology, but they would also bring...working with CRISPR than just designing a repair template that contains your desired edit, and I want to...CRISPR updates on our blog to keep current with the latest and greatest! Addgene resources 3 Tips to Improve...Editing in Human Cells Hands-on tips for repair template design from guest bloggers Dominik Paquet and ...

  18. Hot Plasmids June 2018 - Reverse transcriptase, nanobody, and protein-DNA interaction tools

    Type
    Blog Post
    Published
    June 27, 2018, 1:16 p.m.
    ...continuing to copy the template rather than dissociating).  MarathonRT, isolated from Eubacterium rectale...fields of biology and enable the conversion of RNA templates to DNA but suffer from several important shortcomings...structured HCV genome (~9.6 kb of highly structured template!). MarathonRT also faithfully transcribes RNA ...protein fused with GFP, YFP, mCerulean, or other related fluorophores. Find all of the nanobody toolkit ...biotin. The biotin-labeled proteins can then be isolated and analysed. In this work, the authors specifically...discovery of proteins involved in transcriptional regulation and chromatin structure at your locus of choice...technology, which they call C-BERST (dCas9–APEX2 biotinylation at genomic elements by restricted spatial tagging...

  19. Mentoring for Scientists: I Have a Mentor, What Now?

    Type
    Blog Post
    Published
    March 6, 2014, 7:12 p.m.
    ...mentor. How do you make the most of this new relationship? Consider adding formality and active goal setting... setting to your mentoring relationships, so that you can reap rewards in the form of reaching career ... career path is a great start to a mentoring relationship, but if you don’t take this past the chatting...meetings and setting the direction in a mentoring relationship falls on the shoulders of the Mentee. A Mentor...progress over the course of an ongoing mentoring relationship (in a book or online). Mentee: Make a list of...secret of accountability To make a mentoring relationship really productive, the most important step you...on setting SMART goals using a Development Plan template. A Development Plan or mentoring journal will ...

  20. Prime Editing: Adding Precision and Flexibility to CRISPR Editing

    Type
    Blog Post
    Published
    Jan. 13, 2025, 9:49 p.m.
    ...efficiency of Cas9 cleavage is relatively high while the efficiency of HDR is relatively low, meaning that most...without double-strand breaks (DSBs) or donor DNA templates.  Figure 1: An overview of prime editing...summarize the major ones in Addgene’s repository later in the post. Advantages of prime editing Less constrained...homology-directed repair Homology-directed repair (HDR) stimulated by double strand breaks has been widely used ...increase activity, enhance binding between the template and primer binding site, increase processivity...remake that strand using the edited strand as the template. The PE3 system does just this by including an...PE = prime editor; RTT = reverse transcriptase template; PBS = primer binding site. Image from Doman et...

  21. New and Upcoming Viral Vectors - September 2019

    Type
    Blog Post
    Published
    Sept. 12, 2019, 1 p.m.
    ... AAV Targeting and manipulating specific, genetically-defined neuronal populations with intersectional...calcium sensors and GABA sensors) Calcium sensors The latest calcium sensors from Janelia (jGCaMP7) continue...potential, GCaMP7s is approximately twice as bright, relative to baseline fluorescence, as the GCaMP7f variant...useful for imaging signaling activity in large populations of densely-labeled neurons because background...all our biosensors inventory, and check out our latest vectors below: pGP-AAV-syn-jGCaMP7f-WPRE (104488...Optogenetics continues to be a useful method for manipulating specific neurons. We recently packaged a soma-targeted...specificity of influence measurements by reducing photostimulation of the axons and dendrites of non-targeted...

  22. Hot Plasmids - December 2019 - CRISPR pooled library, LipoGlo, CRISPRa, and Voltage Imaging Tools

    Type
    Blog Post
    Published
    Dec. 10, 2019, 1:58 p.m.
    ...transduced by lentivirus and have been difficult to manipulate genetically. To solve this issue, Sarah Teichmann...disease is driven by atherogenic lipoproteins that accumulate within the vascular wall. ApoB, the main structural...lipoproteins, shuttles lipoproteins through the circulatory systems, but can also lead to the formation of... of ApoB-LPs has been shown to have a strong correlation with cardiovascular disease and indirect measurement...in vivo model to identify drug candidates that modulate ApoB-LP size and concentration, Steven Farber ...that the system is active in E. coli and closely related species, is amenable to use with other bEBP/UAS...and the system may not be adaptable to distantly related species from E. coli. This tool should serve as...

  23. R Bodies: Membrane-Rupturing Microscopic Tools

    Type
    Blog Post
    Published
    April 14, 2016, 2:30 p.m.
    ... can rupture E. coli spheroplasts to release encapsulated proteins. Video by ACS. For a more cartoony ...thousands of times by propagating through an enormous lattice of identical subunits. A brief history of R bodies...coli in the 1980s  (Kanabrocki et al 1986). Much later, it was shown that putting the operon under a strong...mutants that couldn’t extend at a normal pH and later realized that these mutations clustered in a particular...their ability to penetrate other barriers (like vasculature or extracellular matrices) in response to pH ... response to environmental conditions. More speculatively, R bodies might endow divergent organisms with...Protein Piston That Breaks Membranes to Release Encapsulated Cargo. ACS Synth. Biol. (2016). doi:10.1021/...

  24. Why and how to start studying chromatin (with Addgene's help)

    Type
    Blog Post
    Published
    June 12, 2018, 11:40 a.m.
    ...gene a megabase away, or looking at the histone methylation status at a double-strand break that needs repairing...chromosomes interact with each other. Chromatin regulation is complex and influences--directly or indirectly...difficult to consolidate every conceivable chromatin-related plasmid in a collection that grows on a daily basis... Yi Zhang Leonard Zon  You!     Chromatin-related genes with plasmids available from Addgene The ...directly on our website. In-exhaustive Chromatin-related genes with plasmids on Addgene.org AGO1 BRCA1...from Addgene Many articles reporting chromatin-related research describe the production of plasmids that...2018 Feb 13. pii: 1722491115. Addgene blog posts related to chromatin This is not the first Addgene blog...

  25. Hot Plasmids - June 2019 - Optogenetics, Acoustic Reporter Genes, microRNAs, and the CRISPR-Cas9 system CHIME

    Type
    Blog Post
    Published
    June 6, 2019, 12:41 p.m.
    ... in an unregulated fashion and thus a marker for transfection. The other (mKate2) is regulated in the ...Constructs using the photodimerizer system CRY2-CIB1 to regulate botulinum neurotoxin were also created. The depositors...new light on mechanisms of miRNA-mediated gene regulation Article contributed by Michael Lemieux  Listen...microRNAs (miRNAs) are recognized as important regulators of post-transcriptional gene expression across...several years of research, the mechanisms of miRNA regulation have yet to be fully elucidated. Ron Weiss’s ... present, it will repress expression of mKate2 relative to EBFP2. Weiss’s group used these reporters to...define the different cells present in a mixed population, or identify differentiation and disease states...

  26. A Quick Guide to a Career in Software Product Management

    Type
    Blog Post
    Published
    Nov. 24, 2020, 2:15 p.m.
    ...is software product management, and how does it relate to science? In the simplest of terms, a software...more about the stakeholders, the product managers collate and organize that information and form it into ...science backgrounds, most did not. It was my job to translate complex lab operations workflows into a format... help improve their work. Then, I distill and translate that valuable information back to the rest of ...science companies, try to find internships with translatable skills: working with data, working with people...jargon and know the concepts as soon as possible Later in your career:  Show demonstrated skill of people...project management  Bolster your skills in strong relationship-building and influence. You’ll need a large ...

  27. How to Make Friends and Meet People at a Scientific Conference

    Type
    Blog Post
    Published
    Aug. 7, 2014, 1:58 p.m.
    ...no better place for a scientist to make new relationships than at scientific conferences. Conferences ...this is a great ice breaker. Of course, real relationships go further and grow over time, but being interested...fun, memorable meal is a great way to build relationships. Make a list of experimental problems or questions...work – casual socializing is how people build relationships that are memorable. It isn’t always appropriate...overdo it (some of my best studies were designed over late night poker games at Gordon Conferences). You don... of connection. You can also see and offer congratulations on job changes and promotions.  Follow academic...scientists on ResearchGate and/or via Pubmed alerts. Congratulate your connections when they get a new paper published...

  28. Hot Plasmids April 2018 - Protein Degradation, Nanoscopy, FIRE-Cas9, and Yeast Expression Tools

    Type
    Blog Post
    Published
    April 23, 2018, 2 p.m.
    ...ddifferent cell types, including difficult-to-manipulate primary cells, achieving knockdown within 1-2...methods for TRIM-Away delivery. For small cell populations, co-microinjection of TRIM21 protein and the ...the target antibody works well. In larger cell populations, you can deliver these components together using... electroporated cells re-adhered to the culture plate within 4 hr of treatment. In some primary cell lines... CRISPR/Cas9-based system to study epigenetic modulation. While catalytically inactive cas9 (e.g. dCas9...-minute analysis of the kinetics of chromatin regulation and subsequent mathematical modeling. Its reversibility...amount of noise in reporter gene expression by modulating the induction of transcription factors 1 and ...

  29. Visualizing Protein Turnover In Situ

    Type
    Blog Post
    Published
    Jan. 16, 2018, 3:20 p.m.
    ... in neurons reveals a mechanism for regulating its translatability." Science 343.6169 (2014): 419-422....Therefore, I set out to study how protein turnover is regulated in neurons. Protein turnover is the combined ...the abundance of proteins in cells. Homeostatic regulation of protein abundance is essential for normal ...acids. The use of radioactive materials is highly regulated, requires designated work areas and come with ...proper microscope setup we can observe the whole population of one particular protein in neurons or in any... has recently published techniques to follow translation in living single cells. However, before CRISPR-Cas9...synthesized proteins of interest were detected and later visualized by FUNCAT-PLA. Before these methods ...

  30. Human Germline Editing Using CRISPR

    Type
    Blog Post
    Published
    Aug. 10, 2017, 2:19 p.m.
    ..., and electroporated them with an ssODN repair template in induced pluripotent stem cells made from the...fertilization. Since Ma et al. designed a repair template with two synonymous mutations to distinguish it...were able to distinguish between repair from the template vs repair from the maternal MYBPC3 allele. Interestingly...from the single-stranded oligonucleotide (ssODN) template supplied by Ma et al., but instead from the mother...supplied single-stranded oligonucleotide (ssODN) template. Embryos also underwent editing through the NHEJ...NHEJ pathway. Created with BioRender.com. In later characterization studies, the edited embryos proceeded...embryos to undergo editing using the provided ssODN template. In fact, this finding led the authors to title...
Showing: 151 - 180 of 792 results