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Welcome class! Today we're going to explore the strategies that help us modify gene expression at the RNA level. Letβs start with RNA interference, or RNAi. Who can tell me what RNAi does?
Isn't RNAi used to silence specific mRNA?
Exactly! RNA interference employs small interfering RNA or short hairpin RNA to silence target mRNA, preventing translation. Remember, RNAi = silencing! Now, what about antisense RNA? How does that work?
It binds to mRNA to block translation?
That's correct! Antisense RNA effectively prevents protein synthesis by binding to mRNA. Letβs remember: 'Antisense = Block!' Now, letβs jump into CRISPR-Cas13, which is a fascinating topic!
What does CRISPR-Cas13 do?
Great question! CRISPR-Cas13 is an RNA-targeting enzyme used for knocking down or modifying RNA. It's a game-changer in gene regulation strategies. We'll cover more applications of these technologies later!
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Now that we've discussed RNA strategies, let's talk about mRNA vaccines. Why are these significant, particularly in the context of COVID-19?
They teach our cells to produce antigens to fight viruses?
Correct! mRNA vaccines use engineered transcripts to instruct the body to produce specific antigens. This innovative approach employs the principles of transcriptomic engineering and has revolutionized vaccine development. What would be a key takeaway about these strategies?
We can control gene expression and fight diseases!
Exactly! This control over gene expression opens up avenues not only in vaccines but also in therapeutic solutions. Fantastic insights today!
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The section discusses various engineering strategies aimed at modifying gene expression through transcriptomic techniques and enhancing protein functions via proteomic engineering. It details methods such as RNAi, antisense RNA, and CRISPR-Cas13, along with tools for protein engineering, illustrating their significance in research and medical applications.
In the rapidly evolving field of genetic engineering, transcriptomic and proteomic strategies play a crucial role in manipulating gene expression and protein functions. This section delves into the various engineering techniques used for RNA and protein analysis, which include:
The significance of these tools is underscored as they facilitate post-transcriptional control of gene expression. This section serves to illuminate how these various technologies contribute to advancements in fields like therapeutics and functional proteomics.
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Modify protein sequence to alter function
Site-directed mutagenesis is a technique used to intentionally change specific amino acids in a protein sequence. This alteration can affect the protein's shape and, consequently, its function. By modifying particular components of a protein, scientists can study how these changes influence the proteinβs role in biological processes.
Imagine a car's engine where a specific part (like a spark plug) is intentionally altered to see how it affects the car's performance. Similarly, in genetics, changing a part of a protein can help researchers understand its function and how it can be improved or modified for better results.
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Combine proteins (e.g., GFP-tagged enzymes)
Fusion proteins are created by linking two different proteins to form a new one. This combination can serve various purposes, such as simplifying the study of protein interactions or enhancing visualization. For instance, a fluorescent protein like GFP (Green Fluorescent Protein) can be attached to a protein of interest, allowing scientists to track its location within a cell under a microscope.
Think of fusion proteins like a flashlight with a special color filter attached. The flashlight is the protein, and the filter allows you to see it in a new way. Just as the filter enhances your ability to see details, GFP helps scientists visualize and understand where proteins are functioning within cells.
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Facilitate purification and localization
Protein tags like His-tags and FLAG-tags are short sequences of amino acids added to proteins. These tags serve as markers that allow researchers to easily identify and purify the proteins. For example, a His-tag binds to nickel during purification processes, making it easier to separate the protein from other cellular components.
Consider a library where each book has a special sticker on its spine that makes it easy to find. These stickers represent the protein tags. Just as the stickers help you locate the books quickly, protein tags help scientists isolate and study specific proteins in a complex mixture.
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Mimic natural selection to evolve proteins
Directed evolution is a method that simulates the process of natural selection to develop new proteins with desirable traits. Scientists introduce random mutations into a gene and then select for proteins that exhibit improved functions. This technique allows researchers to create proteins with enhanced properties, like increased stability or better activity under certain conditions.
Imagine a gardener who selectively breeds plants to develop a new variety that produces sweeter fruit. By choosing the best plants each season and breeding them, the gardener creates a stronger plant over time. Similarly, directed evolution enables scientists to refine proteins step by step, enhancing their characteristics through iterative selection.
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Key Concepts
RNA interference allows for specific silencing of genes.
Antisense RNA blocks the translation of mRNA.
CRISPR-Cas13 enables precise modification of RNA.
mRNA vaccines utilize engineered transcripts for immunization.
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Use of RNAi in research to silence specific genes.
Application of mRNA vaccines in the COVID-19 pandemic to enhance immune response.
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RNAi silences, shhh, so genes won't chat,
Imagine a librarian (RNAi) who quietly removes books (mRNA) from the shelves so those stories (proteins) never get told, while the antisense librarian puts a cover on books to prevent checks out at all!
Acronym to Remember RNA Control: 'RAP': RNA interference, Antisense RNA, Proteins modified.
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Review the Definitions for terms.
Term: RNA Interference (RNAi)
Definition:
A mechanism to silence target mRNA thus preventing protein synthesis.
Term: Antisense RNA
Definition:
A molecule that binds to complementary mRNA to block its translation.
Term: CRISPRCas13
Definition:
An RNA-targeting system used for knocking down or modifying RNA.
Term: mRNA Vaccines
Definition:
Vaccines that use messenger RNA to instruct cells to produce antigens.