Understand the concept of an expression vector, how it differs from other vectors and what features it contains

Describe how and why you would want to create a fusion protein

• Translational
• Transcriptional

Understand how methylation can protect against most endonucleases (also which ones can't it protect against?)

Understand how regions of homology can allow you to create transgenic organisms

Be able to describe the ways that we can edit genomes (hint - look at the mechanisms and assays below)

For the following mechanisms, understand/describe (to the detail discussed in lecture):

• The sequence features required for the mechanism to take place
• The steps of the mechanism
• The proteins involved at each step
• The outcome(s) of the mechanism

Mechanisms:

• RNAi
• Site directed mutagenesis
• CRISPR/Cas9

For mechanisms as well as the other proteins mentioned in the learning objectives, think about what would happen if you removed their functionality through mutagenesis.

Assays/techniques to be familiar with:

• Affinity purification
• Epitope tagging
• Western blots
• GFP fusion proteins
• Site directed mutagenesis
• In general, knocking out/silencing a gene (transgenics and RNAi)