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We narrowed to 1,452 results for: ina

Showing: 421 - 440 of 1452 results

  1. CRISPR 101: Homology Directed Repair

    Type
    Blog Post
    ...Originally written by Marcy Patrick and Mary Gearing on Mar 12, 2015; updated by Christina Mork, Jul ... Homologous Recombination References: Repair of Strand Breaks by Homologous Recombination. Jasin M and...DNA template used for repair is identical to the original DNA sequence at the DSB, or it can introduce specific... to as the displacement loop (D loop). The recombination intermediates can then resolve to complete the...newly synthesized sequences are retained by the original damaged DNA molecule. Unlike in DSBR, following...The size of the intended edit is the biggest determinant when selecting a type of donor. ssDNA templates...Mutagenesis (2000). PubMed. Multiple Pathways of Recombination Induced by Double-Strand Breaks in Saccharomyces...

  2. Hot Biosensors 2022: Year-End Roundup

    Type
    Blog Post
    ....cell.2022.07.013 pYtags illuminate RTK signaling Receptor tyrosine kinases (RTKs) are a major class ... size (~1500 bp) makes iGECInano suitable for AAV packaging.    Find iGECInano plasmids here!   Matlashov...opening up new research directions that were inaccessible with earlier tools. They also produce more than...sensitivity, and photostability under two-photon illumination than previous GEVIs. Some constructs are also...the phosphorylated state of an immunoreceptor tyrosinase-based activation motif (ITAM). By fusing ITAM...spatiotemporal measurements of receptor tyrosine kinase signaling in living cells. bioRxiv 2022.08.13.503850....503850; doi: 10.1101/2022.08.13.503850.  iGECInano NIR calcium reporter Near-infrared (NIR) fluorescent...

  3. Troubleshooting and Optimizing a Western Blot

    Type
    Blog Post
    ...so it’s better to start over if that happens.  Finally, if you’re seeing a weird issue and can’t figure...ladder compatible with your imaging system; or a combination of the two. There are many commercially available...SDS for your proteins to migrate…so don’t try eliminating one component entirely. If you are trying to ...bit to find a membrane, transfer, and buffer combination that works. Background If you’re seeing odd background...manufacturer. You can also consider using monoclonal or recombinant antibodies, which have much lower lot-to-lot ...point. You can also consider using polyclonal or recombinant multiclonal antibodies to increase overall signal...enough antibody binding sites Polyclonal or recombinant multiclonal antibodies Protein not properly...

  4. Photosensitizer Induced Cell Ablation with FAP-TAP MG-2I-dL5**

    Type
    Blog Post
    ...vivo.  Read on to learn more about this killer illumination technique! What is a photosensitizer? First...species (ROS). The ROS produced by photosensitizers inactivate target proteins or cells while having little ...within 30 minutes of exposure. 90 seconds of illumination were sufficient for 100% effective killing of...the dye. Embryos exposed to just 12 minutes of illumination had fewer mCer3-labeled cells in their hearts...photosensitizer protein that generates ROS when illuminated with green light. They found that KillerRed required...allows for precise light-based control of protein inactivation/cell ablation while having limited collateral...comments below!   References 1. He, J., Wang, Y., Missinato, M. A., Onuoha, E., Perkins, L. A., Watkins, S...

  5. New Neuroscience Tool: The iGluSnFR3 Glutamate Sensor

    Type
    Blog Post
    ...localization through different C-terminal anchoring domains The original iGluSnFR in the pMinDisplay backbone...modified form of the cytosolic C-terminal domain of Stargazin including a terminal PDZ ligand. The researchers...This post was originally written by Tyler Ford in 2018. It was updated by guest blogger Abhi Aggarwal... explore advances that have happened since our original 2018 blog post. The origins of iGluSnFR   iGluSnFR... on the extracellular side. In addition to the original PDGFR TMD, the researchers introduce iGluSnFR3...adapted from Aggarwal et al… Figure 4. PDGFR is a C-terminal fusion to the PDGFR transmembrane domain in the...expression pMinDisplay vector. GPI contains a C-terminal glycosylphostidylinositol anchor. SGZ contains...

  6. What Good Citizenship Can Do for Reproducibility in Science

    Type
    Blog Post
    ..., the problem comes down to inadequate information about what the original researchers actually did. Admittedly... have to be this way. We can never completely eliminate the risk that a minor variation in experimental...put down a basic outline and refer back to the original paper that everyone in your lab has been referencing...President of Editorial at Cell Press.  She trained originally in cell biology, completing her PhD at the MRC...

  7. Celebrate National Book Month with Addgene

    Type
    Blog Post
    ...atypicalities, including a patient with amusia - the inability to perceive music as anything but noise. As a ...pop psychology, Musicophilia is accessible and fascinating, all without compromising on the relevant science... Tsang, Science Communications and Marketing Coordinator Non-science favorites  The Mistborn series by...even more sinister. 1000 pages long. Incredibly imaginative, funny, bittersweet, and shocking at times."     ... PhD, majored in Biochemistry or International Finance. A book club could lead to smart, sometimes contentious...

  8. Improving Transduction Efficiency with Exosome AAVs

    Type
    Blog Post
    ...demonstrated in different cellular environments such as retina or glioblastoma). They also found that exo-AAVs... protects them from antibody recognition and inactivation (György et al., 2014), but scientists have yet...deafness (György et al., 2017), exo-AAV2 in mouse retina after intravitreal injection to transduce bipolar...difficult to reach, and are more resistant to inactivation through anti-AAV antibodies. References György...vector mediates robust gene delivery into the murine retina upon intravitreal injection." Scientific reports...

  9. How to Deposit Your Plasmids with Addgene

    Type
    Blog Post
    ...you to quickly associate your plasmids with the original publication where you described them. Search for...This publication will be listed at the top of the final plasmid pages and scientists will be asked to cite...repetitive sequences (which can be prone to recombination in bacteria) such as lentiviral, retroviral,...Verification On this page you will see a preview of the final plasmid page that will appear on Addgene’s website...available to the research community. This article was originally written by Tyler Ford in January 2016. It was...was updated by Angela Holmes and Christina Mork in February 2024.  Additional Inside Addgene Content on...

  10. Antibodies 101: Multiplex Immunofluorescence

    Type
    Blog Post
    ... one target is labeled and imaged at a time, inactivating the fluorescent secondary antibodies before ...comes in. After you image the first target, you inactivate the fluorophores in the sample, either by chemical...new primary, wash, add secondary, image again, inactivate, and repeat these cycles for all your targets...from. But by labeling sequentially with one, inactivating the fluorophores, then labeling with the other...brain samples from the James Trimmer Lab, using recombinant antibodies with different isotypes. (A) Neocortex...antibodies are commercially available. Pick whichever combination of host and target species, isotype specificity... 2A above, Andrews et al. used the following combination of antibodies:   Target (Rat) Primary antibody...

  11. PiggyBac-ing Through the Genome Editing Field

    Type
    Blog Post
    ...piggyBac™ transposon? The piggyBac™ transposon was originally identified in moths in 1983, but wasn’t successfully...or when the gene is constitutively-active or a dominant-negative mutant. Reducing the copy number can ...host machinery for DNA repair by homologous recombination, which results in the incorporation of your ...containing a negative selection marker such as thymidine kinase, a researcher may subsequently kill off any cells...site-specific nuclease to stimulate homologous recombination and Footprint-Free™ removal of the selection...First, instead of using piggyBac™, you could use recombinase technology (Cre/loxP) to remove the donor cassette...[7], it is generally not as efficient as Cre recombinase. However, the use of negative selection largely...

  12. GCE4All: Making Genetic Code Expansion Accessible

    Type
    Blog Post
    ... the synthetic biology scene in 1989 through a seminal paper from Peter Schultz’s lab describing a technique...same for all three components, meaning that the final protocol may involve a sensitive balance of trade-offs...GCE4All is a great place to start. They host monthly webinars, workshops, and a bulletin board on their website...their first protocol is available now. It is for recombinant expression of phosphoserine-including proteins...Site-specific Incorporation of Phosphoserine into Recombinant Proteins in Escherichia coli. 12(21). https:/...

  13. Antibodies 101: ChIP

    Type
    Blog Post
    ...washing and then incubate with Proteinase K to reverse the crosslinking. Finally, purify your DNA with a PCR...fragment size and (2)  incubate with RNAse and Proteinase K to purify the DNA and then determine concentration...isolated your antibody:protein:DNA complexes and finally (5) isolated the DNA from your antibody:protein... on Addgene.org Find plasmids encoding recombinant antibodies at Addgene ...

  14. Opto-Nanobodies: Using Light to Manipulate Cell Signaling and Protein Purification

    Type
    Blog Post
    ...their target protein is altered by blue light illumination. This ability to reversibly bind and release...tagged, which limits these approaches to use with recombinant proteins. Purifying an untagged protein with ...initiates the extracellular signaling-regulated kinase (ERK) signaling cascade. This signaling is a lot...signaling by using a fluorescent reporter for Erk kinase translocation. These results demonstrate that OptoNBs...pathway. What’s next for OptoNBs? While OptoNBs illuminate a path to light-controlled protein binding, they...

  15. Plasmids 101: Inducible Promoters

    Type
    Blog Post
    ...expression and largely defines the amount of recombinant protein that will be produced. Many common promoters...Positive inducible In the OFF state, the promoter is inactive because the activator protein cannot bind. After...Negative inducible In the OFF state, the promoter is inactive because a bound repressor protein actively prevents...tetracycline-controlled transactivator) is normally inactive and cannot bind the tetracycline response elements...a reporter gene. When light is present, YFI is inactive, preventing repressor cI synthesis and allowing...

  16. Teaching an Old DOG New Tricks: Controlling Protein Activity with GFP

    Type
    Blog Post
    ...another biological workhorse, Cre recombinase, which induces recombination at LoxP sites. Placing a lox-STOP-lox...researchers to activate transcription or Cre recombinase activity only in the presence of GFP. These systems... transcription. Thus, T-DDOGs can be used in combination with green-red Cre-lox systems. T-DDOGs can also...T-DDOGs, GFP, and a reporter construct into the mouse retina. GFP expression robustly activated the reporter...Cre-DOG is both robust and specific. When tested in retinal electroporation studies, DsRed reporter expression...system, including the motor cortex, cerebellum, and spinal cord. Cre-DOG is also suitable for optogenetic ...type-specific manipulation with GFP-dependent Cre recombinase.” Nat. Neurosci. 18(9) (2015): 1334-1341. PubMed...

  17. 6 Tips for Grant Writing

    Type
    Blog Post
    ...novel way. Finally, engineering new applications to solve problems will provide originality to your work...on how to submit your work either through seminars or webinars. Sending on your grant application to your... the deadline isn’t going to be much use. 2. Originality is essential for grant success The two most important...be looking for in your research proposal are originality and relevance. When reading through your proposal...our own or other groups, but your work must be original (otherwise why would you be interested in it?)...Frontier Science Program's website. One way to add originality to your work is to collaborate with specialists...your work, or even the successful use of your preliminary data to secure further funding. Make it clear...

  18. Important Considerations When Using AAVs

    Type
    Blog Post
    ...communities. Many thanks to our guest blogger Katrina Armstrong. Katrina Armstrong is a Neurophysiology Msc Student...This post was contributed by guest blogger Katrina Armstrong, a Neurophysiology Msc Student at the University...which cell type? Location (3): Traveling from the original location - Axonal Transport in the CNS What tissue... Photoreceptor Cells AAV2, AAV5, AAV8 RPE (Retinal Pigment Epithelium) AAV1, AAV2, AAV4, AAV5, AAV8...your particular experiment! Traveling from the Original Location - Axonal Transport It has been demonstrated...serotonin in movement. Follow her on twitter @katrinaboards.   References 1. Russell, David W. "AAV vectors...and Richard J. Samulski. "Self-complementary recombinant adeno-associated virus (scAAV) vectors promote...

  19. A History of Genome Engineering in Popular Culture

    Type
    Blog Post
    ... that exterminate entire humanity were the two most popular topics that sparked the imagination of most... CBE but use a ssDNA adenosine deaminase (evolved from a tRNA deaminase enzyme, TadA) to induce a A→G ...hybrids. This book is published 30 years after the seminal work of Gregor Mendel on the genetic inheritance... first ever X-ray diffracted image of the DNA. Finally, in 1953, Franklin along with James Watson and ...Thurman and Jude Law. The movie’s name is itself a combination of the four bases of DNA – A, T, G, and C. In...These breaks can then be repaired by homologous recombination with a repair template.  Each zinc finger recognizes...The cytidine base editors (CBE) use a cytidine deaminase to introduce a C→T change at precise locations...

  20. Click Biology: How to Click and Play to Explore Biology

    Type
    Blog Post
    ... Cysteine-less, fast split intein; N-terminal CLm and C-terminal Aes gp41-1 Split inteins A small, fast...interfaces and the freedom to test out lots of combinations, to end up finding something surprisingly useful...attB2r-SNAP-attB3 SNAP-tag Gateway entry clone to add C-terminal SNAP-tag Promega Plasmid Collection HaloTag ...PCV-Cas9, Cas9-PCV HUH-tag Cas9 plasmids with N- or C-terminal PCV2 HUH-tag; increase homology-directed repair...
Showing: 421 - 440 of 1452 results