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  • Rat Brain Case Study

    1099 Words  | 5 Pages

    1000 µg rat brain lysate was incubated with 50 µg construct overnight on a shaking incubator at 4°C. Samples were loaded on a polyacrylamide gel and processed for immunoblotting (see appendix) Immunoblotting for sacsin and JIP3 interaction Primary Sacsin and JIP3 antibodies (Santa Cruz cat. sc-515118 and sc-46663) raised in mice were added in a 1:3000 and 1:2000 ratio respectively to the corresponding membrane sections and incubated on a shaking incubator for 1 hour. Next, excess antibody was removed

  • Dysbindin-1 Case Study

    954 Words  | 4 Pages

    dysbindin-1 binds to Drp1 to enhance its oligomerization. To test this possibility, we transfected HEK-293 cells with YFP-Drp1 along with HA-tagged dysbindin-1C or -1A, and used an anti-HA antibody to immunoprecipitate dysbindin-1 at 2 days after transfection. Drp1 was detected in the immunoprecipitation product from cells transfected with dysbindin-1C, but not with -1A plasmid (Figure 4G), indicating that

  • Mir34a Essay

    1457 Words  | 6 Pages

    capability of Aβ clearance in AD, we suspected that miR34a may be an indispensable player. To validate this, Aβ42 fibrils were prepared from Aβ42 monomer ex vivo and used to treat glioma A172 and U251 cells. Aβ uptake was blocked by both transient transfection of a miR34a mimic and stable miR34a overexpression (Supplementary Fig. 1B, Fig. 1A), but enhanced by anti-miR34a (Fig. 1B), indicating that miR34a suppressed Aβ phagocytosis in these cells. The fact that enhanced and decreased levels of miR34a

  • T Cell Gene Therapy : The Center Of Research With Stem Cell Transfection In Geney Cells

    941 Words  | 4 Pages

    The CCR5 co-receptor is not only the center of research with stem cell transplant, but has also been a recent target of gene therapy research. Gene therapy is a fairly new technology where genes (edited or normal) are transplanted in humans to produce a specific response. One avenue of research that is currently being studied is T cell gene editing focused on the CCR5 delta32 mutation in HIV infected patients using the CRISPR/cas9 system. There is some research using TALENS, but evidence with CRISPR/cas9

  • Ultrasound Gene Delivery

    680 Words  | 3 Pages

    cavitation bubbles will disruptive the membrane of cells and tissues to enhance drug transport. Use the ultrasound to enhance cell permeability is a new methods for gene transfection. The increase uptake of DNA or RNA has been demonstrated by several investigators2-1, 2-5. However, the further enhancement of gene transfection

  • Dendrimers Research Paper

    297 Words  | 2 Pages

    dendrimers are the most frequently used due to their high transfection efficiency. The transfection efficiency is highly influenced by the size of dendrimers and the type of amine groups that they are composed of. The primary amine groups participate in DNA binding, so they promote DNA cellular uptake. However, the tertiary amino groups reinforce the release of DNA into the cytoplasm because they act as a proto-sponge in endosomes. Transfection efficiency can be enhanced by increasing the size and diameter

  • The Rna Interference ( Rnai ) Pathway

    2224 Words  | 9 Pages

    The RNA interference (RNAi) pathway is an important biological system that is utilised by many organisms as a method of regulating gene expression. But what exactly is RNAi? And what is the importance of its use and application in biology? RNAi is a cellular process that actively silences specific genes. This method of post-translational gene regulation has been observed in many organisms including plants, fungi, some bacteria and animals. RNAi inhibits protein synthesis of the target transcript

  • Western Blotting Of Camkii Œí And T 287

    847 Words  | 4 Pages

    Western blotting of CaMKIIβ and T-287 phosphorylated CaMKIIβ Western blotting was performed as described previously (Chapter 3). In short, Proteins were extracted from OLs using RIPA buffer with protease and phosphatase inhibitors. The concentration of extracted proteins was assessed using the BCA assay. Target proteins were probed with primary antibodies for CaMKIIβ (1:1000, Life Technologies) and phosphor-CaMKIIβ (T287) (1:1000, Abcam) at 4°C overnight. Membranes were incubated with appropriate

  • Lab Report

    1214 Words  | 5 Pages

    lung cancer cells, we used real-time PCR to measure the expression of miR-365b-3p in various cells. Results showed that the expression of miR-365b-3p was significantly lower in lung cancer cells compared with that in normal cells (Figure 1). After transfection, the expression of miR-365b-3p in COLO 668 cell line was successfully upregulated by miR-365b-3p mimic and inhibited by miR-365b-3p inhibitor (Figure 2A). Cell viability assay showed that miR-365b-3p suppressed cell viability while miR-365b-3p

  • Human, Eukaryotic And Vitro Associations Of Murine Sec Specific Ef ( Eefsec )

    976 Words  | 4 Pages

    INTRODUCTION In eukaryotes, Usually the UGA acts as a stop codon in transcription but the elongation factor, known as SELB, has a C-terminal SECIS RNA binding domain and N-terminal Sec-tRNA domain, inserts the selenocystiene at the stop codon. An SECIS binding protein, known as SBP2 is found in mammals but it lacks EF functionality. We will describe the in vivo and in vitro associations of murine SEC-specific EF (eEFSec) element with SBP2 and SECIS in order to correlate their function in selenoprotein