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This homework is based off of the Week 7 Lab. This is a good week to start honing in final projects and focusing on developing / researching protocol.

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<aside> <img src="/icons/push-pin_green.svg" alt="/icons/push-pin_green.svg" width="40px" /> Key Links: https://docs.google.com/document/d/1DwQ7I2By4BIKbnY48m6iQ_081TFI-C4xmoVOrPgcNVQ/edit?tab=t.0

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Questions 1-3 are mandatory for all students.

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  1. How do endoribonucleases (ERNs) work to decrease protein levels?

Endoribonucleases (ERNs) cleave specific mRNA molecules inside the cell. By cutting the mRNA, they prevent it from being translated into protein, reducing the amount of protein produced.

Name 2 differences between how ERNs work and how proteases work.

Feature Endoribonucleases (ERNs) Proteases
Target Cut mRNA (genetic message) Cut proteins (finished product)
Stage of action Prevents protein production (early, at the RNA level) Destroys existing proteins (late, after proteins are already made)
  1. How does lipofectamine 3000 work?

→ Lipofectamine 3000 helps deliver plasmid DNA into human cells by forming lipid-DNA complexes.

These complexes fuse with the cell membrane and allow the DNA to enter the cell, where it can reach the nucleus and be expressed.

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Step-by-step simplified:

  1. Lipofectamine 3000 forms tiny lipid bubbles around the plasmid DNA.
  2. These bubbles merge with the cell’s lipid membrane.
  3. The DNA is released inside the cell, where it can be transcribed and translated into proteins.

How does DNA get into human cells and how is it expressed?

DNA is introduced into human cells through a process called transfection.

In this lab, Lipofectamine 3000 is used to: