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We narrowed to 922 results for: tro

Showing: 381 - 400 of 922 results

  1. Antibodies 101: Conjugation

    Type
    Blog Post
    ...fluorescence microscopy, flow cytometry, or spectrophotometry. The range of fluorophores available for antibody...capable of producing a visible signal, but it has a strong affinity for avidin and streptavidin, both of which...modifying the antibody sequence or that rely on strong protein-protein interactions with other peptides...of non-canonical amino acids or peptide tags. 5) Strong protein-protein interactions with other peptides...of antibody conjugation. But hopefully, this introduction provides a good base from which to dive deeper...

  2. Working with Nuclear Receptors

    Type
    Blog Post
    ...reproduction. Both physiological processes are controlled by deliberate and compulsory actions instigated...corticosteroids, progesterone, androgens, and estrogens), fat-soluble vitamins A and D, thyroid hormone...complex can serve as a rate-limiting conduit to control the actions of the whole complex.  For example,...adipogenesis and macrophage activation. Our mass spectrometry-affinity purification proteomic profiling of...N. J.and O'Malley, B. W. (2002). Combinatorial control of gene expression by nuclear receptors and coregulators...

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

    Type
    Blog Post
    ...derived from the E. coli Ec48 retron RT and the S. pombe Tf1 retrotransposon RT, respectively. Their small...less than 10%. In contrast, the original paper introducing prime editing demonstrated ~20-50% efficiency...fold (Anzalone et al., 2019). PE3, as originally introduced, relies on the PE2 enzyme. PE4 and PE5: Engineering...ligases to act first. PE4 and PE5, as originally introduced, rely on the PE2 enzyme. PEmax: Optimizing prime...found that the mutations that emerged depended strongly on the target edit (Doman et al., 2023). Therefore...

  4. Pathways Over Time Plasmids Engage Students in Functional Genomics Research

    Type
    Blog Post
    ...could provide the basis for student projects in introductory labs. Genome projects use computational methods... of MET/Met genes from the pYES2.1 plasmid is controlled by the GAL1 promoter. The cloning procedure adds...absence of methionine. Since the GAL1 promoter controls MET/Met gene expression, complementation should...PMC232622. 6. Winzeler, D.A., Shoemaker, D.D., Astromoff, A., Liang, H., Anderson, K., Andre, B., Bnangham...over time: Functional genomics research in an introductory laboratory course. CBE-Life Sci. Educ. 17, ar1...

  5. Adeno Associated Virus (AAV) for Cell and Gene Therapy

    Type
    Blog Post
    ... disease. To do so, their T-cells were grown in vitro, their mutations corrected, and the T-cells were... body and thereby correct the mutations using retroviral vectors. However, many diseases require techniques... B, LPL deficiency, Cystic fibrosis, Muscular Dystrophy, Parkinson’s disease, and HIV. A notable landmark...infect target tissues. Improved specificity through control of gene expression and the selection of specific...CRISPR-AAV vectors Learn about Addgene AAV Quality Control Process Resources on Addgene.org Find ready-to-use...

  6. Antibodies 101: Stripping and Reprobing Western Blots

    Type
    Blog Post
    ...better than a nitrocellulose membrane. If you have already run your blot using nitrocellulose, you may decide...isn’t too different, consider antibody affinity: stronger binding antibodies will require harsher stripping...instructions for the membrane you are using. Dry nitrocellulose membranes are brittle, so proceed with caution...depend on your blot — antibodies that bind more strongly or that have saturated the blot may need longer...

  7. Hot Plasmids - October 2020

    Type
    Blog Post
    ...Tsang Fluorescent proteins (FPs) traditionally introduced in Caenorhabditis elegans on extrachromosomal...gene into the target gene’s native locus, but introducing large genetic changes in C. elegans with CRISPR... donors (ssODN). So how can you use CRISPR to introduce large FP genes into C. elegans? By using a split...protein. They used CRISPR with a ssODN template to introduce the small wrmScarlet11 which is less than 60 nt...exclusively in huge bacteriophage genomes. It’s active in vitro and in human and plant cells. Its small size offers...

  8. Tips for Titering Your Lentiviral Preps

    Type
    Blog Post
    ...method can be the absence of suitable controls. The best control for this type of assay is a clonal cell...using a reverse transcriptase mutant in a separate control reaction. This will allow you to quantify the amount...of the qPCR target gene. Developing the proper control line typically requires transducing with a limiting.... 3. Zhang, Bing, et al. "The significance of controlled conditions in lentiviral vector titration and...

  9. Hot Plasmids February 2024

    Type
    Blog Post
    ...systemic AAVs with endothelial-specific tropism in rodents and broad tropism in primates. Nature Communications...Peptide-assisted genome editing for primary cells By Susanna Stroik Even with the advances in CRISPR technology over...difficult to genetically edit due to toxicity from electroporation or poor cellular entry with other transfection...viral vector has brain endothelial cell-specific tropism, with slightly reduced efficacy compared to AA9...

  10. Experimenting in an Open Source lab: from CRISPR to Cats

    Type
    Blog Post
    ... our members took an Arduino microcontroller and a motor controller and made it spin again!  We’ve also...scientists can get involved in modelling, the electronics folks can build a robot worm on top of that model...instance, the centrifuge in the picture had a broken controller and would not spin. It was going to cost far ...far more than it was worth to repair the controller and a new replacement was not possible, but one of our...

  11. Adenoviral Delivery of CRISPR/Cas9 Aims to Expand Genome Editing to Primary Cells

    Type
    Blog Post
    ... mutated gene. In a Scientific Reports paper introducing the delivery method in 2014, Manuel Goncalves...include their episomal nature and very efficient introduction of DNA into therapeutically relevant, non-transformed...B-thalassemia, Sickle Cells Disease, Duchenne Muscular Dystrophy, Alpha1-antitrypsin deficiency (most widely used... for cancer therapy. Elsevier, pp 39–67 Ding Q, Strong A, Patel KM, Ng S-L, Gosis BS, Regan SN, Cowan ...adenoviral delivery of CRISPR/Cas9. Journal of Controlled Release 298:128–141 . https://doi.org/10.1016...

  12. PCR Applications for Copy Number Variation Assays

    Type
    Blog Post
    ...qPCR assays, samples, standard curves, and normal controls must be run in triplicate.  The nice thing about... of DNA in your sample relative to your normal control - you can set up your calculations so that your...so forth. This also serves as a light technical control - an RQ value of 1.5 means you have either made... does not require a standard curve or a normal control, it can take a few hours longer than qPCR to run...AT-rich DNA and by variation in amplicon length. Am J Trop Med Hyg. 2008 Dec;79(6):887-92. PMID: 19052298. ...

  13. Addgene's Cancer Collection Pages Connect the Research Community

    Type
    Blog Post
    ...same result – uncontrolled movement of the vehicle or, in the case of cancer, uncontrolled cell proliferation... cellular level there is a universal theme - uncontrolled cellular proliferation. We have appreciated ...appreciated for some time the causes of this uncontrolled proliferation, in particular the roles of proto-oncogenes...opposite task, keeping cells from dividing out of control. One may compare the roles of proto-oncogenes and...

  14. Google Forums Round Up: First Impressions of NgAgo

    Type
    Blog Post
    ...of all distributed plasmids as part of quality control, but does not perform functional tests. The newest...produced from a plasmid in cells. Additionally, in vitro assembly of NgAgo/ssDNA requires incubation at 55...can conveniently transfect/inject cells with in vitro preassembled Cas9/gRNA complexes or with guide RNA...does however rely on cloning into a vector or in vitro synthesis, which could be less desirable if one ...et al. "DNA-guided genome editing using the Natronobacterium gregoryi Argonaute." Nature biotechnology ...

  15. Genome-wide Screening Using CRISPR

    Type
    Blog Post
    ...drug-resistant population compared to a non-treated control group. In this scenario, gRNAs that are “enriched...future therapeutic targets for diseases causing uncontrolled cell growth, such as cancer. Figure 2...As with any experiment, the use of appropriate controls, multiple replicates and several cell types can...of multiple gRNAs targeting the same gene can be strong evidence that a particular gene is actually important...Titering Your Lentivirus Plasmids 101: NGS Quality Control for Pooled Libraries Find More CRISPR Resources...

  16. Starter guide to induced pluripotent stem cells (iPSCs) part 1:  A renaissance in regenerative medicine

    Type
    Blog Post
    ...preimplantation embryos that are produced during in vitro fertilization treatments and donated afterwards ... produced Ethical concern High. Embryos are destroyed during ESC isolation. Low Chances of post-transplantation... the reprogramming factors were delivered via retroviral infection [8]. The work was taken forward the...generation of human iPSCs from dermal fibroblasts via retroviral and lentiviral infections respectively [9, 10...cell types. Like ESCs, iPSCs can differentiate in vitro into derivatives of all three germ layers, form ...

  17. Simplify Cloning with in vivo Assembly

    Type
    Blog Post
    ...techniques involve the assembly of a circular plasmid in vitro, before transforming it into E. coli for propagation...specifically cuts methylated DNA, so it will selectively destroy the template, but not newly synthesized sequences...protein-coding sequence, or a mutation can be introduced in a gene, while adding a GFP-tag (Figure 3)....unamplifiable’ Check your amplification by gel electrophoresis. In our experience, good amplification after...interested in neuronal signal transduction, and uses electron cryo-microscopy to understand receptor function...

  18. AAVs in Retinal Gene Therapy

    Type
    Blog Post
    ...diseases such as hemophilia, muscular dystrophy, spinal muscular atrophy, neurodegenerative diseases and metabolic...life-threatening (cystic fibrosis, hemophilia, muscular dystrophy)? AAV gene therapy started in the mid-90's with...larger transgenes. Unfortunately it has limited tropism in the retina, primarily infecting cells of the...epithelium (RPE). AAVs, on the other hand, have broad tropism in the eye. AAV-2, the first identified serotype...

  19. MXS Chaining

    Type
    Blog Post
    ... create a single polycistronic 15 kb insert. The final construct was then introduced to HeLa cells (Figure...resulting HeLa cells show robust labeling, with strong expression and detection of each fluorophore at...meaning that (in the previous example) one can control whether the third module is added to the 5’ terminus...degree of identity, so not suited to generate polycistronic containing multiple identical polyadenylation...

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

    Type
    Blog Post
    ...generated a series of retroviral vectors expressing tagged dCas9 and guide RNA. The retroviral enChIP system ...requested plasmids for enChIP through Addgene. Retroviral Expression System for enChIP using CRISPR Addgene...Addgene: Your lab recently developed a retroviral enChIP system using CRISPR technology. Can you describe...Associated with an IFNgamma-Responsive Promoter by a Retroviral Expression System for enChIP Using CRISPR. Fujita...
Showing: 381 - 400 of 922 results