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What happens when DNA double strand breaks occur
Cell activates DNA repair pathways
What are the two major DNA repair pathways
HDR and NHEJ
What is HDR
Precise DNA repair using homologous template
When does HDR occur
S phase of cell cycle
What template is used for HDR
Sister chromatid or donor template
Why is HDR used for knock in studies
Allows precise insertion or correction
How long must homology arms be
More than 60 base pairs
Why are PAM sites removed from HDR templates
Prevent repeated cutting
What is NHEJ
Error prone DNA repair pathway
What type of mutations does NHEJ introduce
Insertions and deletions
What are indels
Insertions or deletions in DNA
Why does NHEJ disrupt gene function
Frameshifts or stop codons
Why is NHEJ used for knockouts
Efficiently inactivates genes