Please help me with the Cellular Biology question below. It is one question, the data given is to help answer it. Please be as detailed as possible. **Investigation of miR-145 Role in Gastric Cancer****Experimental Approach:**1. **Objective:** - To examine whether microRNA-145 (miR-145) influences gastric cancer by comparing levels of Ets1 and miR-145 in both cancerous and normal adjacent tissues from 84 patients using qPCR. 2. **Specific Steps:** - **(A)** Measure the levels of miR-145 and Ets1 in cancerous and corresponding normal tissues. - **(B)** Compare these levels in normal (GES-1) vs. cancer cell lines. - **(C)** Transfect cancer cell lines with Ets1 3’-UTR reporter plasmid and either miR-145 or a control, and then perform luciferase assays. - **(D)** Perform Western blots for cells transfected with miR-145 or mock to detect Ets1 presence.**Results:****A. Relative Levels of Ets1 and miR-145 in Tissues (Figure A):**1. **Ets1 Expression:** - The box plot on the left shows a significant increase in Ets1 transcript levels in cancer tissues compared to normal adjacent tissues. - p < 0.0001 indicates a highly significant difference.2. **miR-145 Expression:** - The box plot on the right demonstrates a significant decrease in miR-145 expression in cancer tissues compared to normal tissues. - p < 0.0001 also indicates a highly significant difference.**B. Relative Transcript Levels in Cell Lines (Figure B):** - Bar graph presenting Ets1 and miR-145 expression levels across different cell lines (GES-1, AGS, SGC-7901, MKN-45). - miR-145 and Ets1 expression levels vary significantly with lower miR-145 and higher Ets1 in cancer cell lines compared to GES-1 cells. - '*' indicates statistical significance.**C. Luciferase Assay with Ets1 3’-UTR Reporter Plasmid (Figure C):** - Schematic diagram outlining the constructs used for the luciferase assay with wild-type (WT) and mutant (Mut) Ets1 3’-UTR sequences. - The sequences provided for both WT and Mut Ets1 3’-UTR

A short RNA molecule called a microRNA, or miR-145 is involved in the post-transcriptional…

Refer to the experimental approach (i.e., very briefly describe) and give the results AND conclusions. The answer is incomplete without reference to all the data including controls. (A) To investigate whether the microRNA-145 (miR-145) plays a role in gastric cancer, levels of Ets1 and miR-145 in cancer and normal adjacent tissue of 84 patients were determined by qPCR. (B) Levels in normal (GES-1) and cancer cell lines were determined. (C) Cancer cell lines were transfected with the Ets1 3'-UTR reporter plasmid and either miR-145 or negative control (mock), and luciferase assays were carried out. (D) Western blots of cells transfected with miR-145 or mock stained for Etsl

Refer to the experimental approach (i.e., very briefly describe) and give the results AND conclusions. The answer is incomplete without reference to all the data including controls. (A) To investigate whether the microRNA-145 (miR-145) plays a role in gastric cancer, levels of Ets1 and miR-145 in cancer and normal adjacent tissue of 84 patients were determined by qPCR. (B) Levels in normal (GES-1) and cancer cell lines were determined. (C) Cancer cell lines were transfected with the Ets1 3'-UTR reporter plasmid and either miR-145 or negative control (mock), and luciferase assays were carried out. (D) Western blots of cells transfected with miR-145 or mock stained for Etsl

Biology 2e

2nd Edition

ISBN:9781947172517

Author:Matthew Douglas, Jung Choi, Mary Ann Clark

Publisher:Matthew Douglas, Jung Choi, Mary Ann Clark

Chapter16: Gene Expression

Section: Chapter Questions

Problem 33CTQ: Describe how RBPs can prevent miRNAs from degrading an RNA molecule.

See similar textbooks

Similar questions

True or False. 1. a.) RNA polymerase decodes mRNA so the ribosome can make proteins. b.) Only coding RNA can interact with the ribosome. c.) The ribosome is composed of both protein and ncRNA. d.)The ncRNA components of the ribosome behave as a ribozyme. Pick one of the FALSE statements from the 4 previous questions and explain why it is incorrect.
Give typing answer with explanation and conclusion a) List three eukaryotic gene expression mechanisms that do not occur in prokaryotes. For two of these, give specific examples and the functional outcomes. b) Describe what is meant by the term “RNA silencing”. c) Using diagrams, give two examples of RNA silencing mechanisms and indicate one difference.
E22. The method of Northern blotting is used to determine the amount and size of a particular RNA transcribed in a given cell type. Alternative splicing (discussed in Chapter 12) produces mRNAs of different lengths from the same gene. The Northern blot shown here was made using a DNA probe that is complementary to the MRNA encoded by a particular gene. The mRNA in lanes 1 through 4 was isolated from different cell types, and equal amounts of total cellular MRNA were added to each lane. 2 3 4 Lane 1: MRNA isolated from nerve cells Lane 2: MRNA isolated from kidney cells Lane 3: MRNA isolated from spleen cells Lane 4: MRNA isolated from muscle cells Explain these results. | |
Diagram the fate of the dsRNA once it enters a cell, highlighting the enzymatic reactions that eventually result in down-regulation of gene function. What is the hypothesised molecular basis of the hypersensitivity to RNAi in the eri-1/lin15b mutants?
Q. When dsRNA is treated with Dicer enzyme siRNA and miRNA’s are produced. What role they will play in gene regulation and expression and how? (Subject Bioinformatics)
What is the difference between crRNA and tracrRNA? Why are both needed for Cas9 to function? What does PAM stand for? Where is it found? What is the difference between Non-homologous End Joining (NHEJ) and Homology Directed Repair (HDR)?
Shown is part of the coding strand of the HBB gene involved in sickle cell disease. What will be the effect on the resulting peptide of a deletion of the two nucleotides shown in bold/underline? 5' CTG ACT CCT GAG GAC 3' a) A change from glutamic acid to valine will occur b) The deletion will not result in a change to the polypeptide c) A nonsense mutation will occur d) The polypeptide will be missing one amino acid, but the reading frame will be preserved
Explain how the data of Fire and Mello suggested that doublestrandedRNA is responsible for the silencing of mex-3 mRNA.
Propose some specific uses of a modified CRISPR-Cas system as a general RNA-guided device for altering cellular functions. What might these functions be, and how could CRISPR-Cas be used to study them?
What would be a possible result if a miRNA that perfectly complemented E3 ligase was successfully processed in the cell?
MAKE CONNECTIONS Compare the CRISPR-Cas systemto the miRNA system discussed in Concept 18.3, includingtheir mechanisms and their functions.
List five types of cancer in which ncRNAs can be involved.