Identify and describe the steps that lead to the secretion of proteins from eukaryotic cells.
Table of contents
- 1. Introduction to Genetics51m
- 2. Mendel's Laws of Inheritance3h 37m
- 3. Extensions to Mendelian Inheritance2h 41m
- 4. Genetic Mapping and Linkage2h 28m
- 5. Genetics of Bacteria and Viruses1h 21m
- 6. Chromosomal Variation1h 48m
- 7. DNA and Chromosome Structure56m
- 8. DNA Replication1h 10m
- 9. Mitosis and Meiosis1h 34m
- 10. Transcription1h 0m
- 11. Translation58m
- 12. Gene Regulation in Prokaryotes1h 19m
- 13. Gene Regulation in Eukaryotes44m
- 14. Genetic Control of Development44m
- 15. Genomes and Genomics1h 50m
- 16. Transposable Elements47m
- 17. Mutation, Repair, and Recombination1h 6m
- 18. Molecular Genetic Tools19m
- 19. Cancer Genetics29m
- 20. Quantitative Genetics1h 26m
- 21. Population Genetics50m
- 22. Evolutionary Genetics29m
11. Translation
Translation
Problem 36c
Textbook Question
The flow of genetic information from DNA to protein is mediated by messenger RNA. If you introduce short DNA strands (called antisense oligonucleotides) that are complementary to mRNAs, hydrogen bonding may occur and 'label' the DNA/RNA hybrid for ribonuclease-H degradation of the RNA. One study [Lloyd et al. (2001). Nucl. Acids Res. 29:3664–3673] compared the effect of different-length antisense oligonucleotides upon ribonuclease-H–mediated degradation of tumor necrosis factor (TNFα) mRNA. TNFα exhibits antitumor and pro-inflammatory activities. The following graph indicates the efficacy of various-sized antisense oligonucleotides in causing ribonuclease-H cleavage. What factors other than oligonucleotide length are likely to influence antisense efficacy in vivo?

1
Consider the stability of the antisense oligonucleotide in the cellular environment. Factors such as chemical modifications (e.g., phosphorothioate backbones or 2'-O-methyl modifications) can enhance resistance to nucleases and improve efficacy.
Evaluate the binding affinity of the antisense oligonucleotide to the target mRNA. This is influenced by the sequence complementarity and the melting temperature (Tm) of the hybridized DNA/RNA duplex.
Account for the accessibility of the target mRNA sequence. Secondary structures in the mRNA or binding of proteins to the mRNA can hinder the hybridization of the antisense oligonucleotide.
Consider the intracellular delivery and uptake of the antisense oligonucleotide. Efficient delivery systems, such as lipid nanoparticles or conjugation with cell-penetrating peptides, can enhance cellular uptake and target accessibility.
Examine the potential for off-target effects. The specificity of the antisense oligonucleotide to the target mRNA sequence is critical to minimize unintended interactions with non-target RNAs, which could lead to side effects or reduced efficacy.

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Key Concepts
Here are the essential concepts you must grasp in order to answer the question correctly.
Antisense Oligonucleotides
Antisense oligonucleotides (ASOs) are short, synthetic strands of nucleic acids designed to bind to specific mRNA sequences. By hybridizing with their complementary mRNA, ASOs can inhibit translation or promote degradation of the target mRNA, thereby regulating gene expression. Their design, including length and sequence specificity, is crucial for their effectiveness in modulating gene activity.
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Ribonuclease H (RNase H)
Ribonuclease H is an enzyme that degrades the RNA strand of RNA-DNA hybrids. In the context of antisense oligonucleotides, RNase H plays a pivotal role in the mechanism of action by cleaving the mRNA once it is bound by the ASO. The efficiency of RNase H activity can be influenced by factors such as the stability of the RNA-DNA hybrid and the presence of other cellular components.
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In Vivo Efficacy Factors
In vivo efficacy of antisense oligonucleotides can be influenced by several factors beyond their length, including cellular uptake, stability in biological fluids, off-target effects, and the presence of nucleases. Additionally, the delivery method and the biological context, such as the type of tissue or disease state, can significantly impact the effectiveness of ASOs in achieving their therapeutic goals.
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