Functional Genomics and Genome Engineering Flashcards
(21 cards)
What is the goal of functional genomics?
Understanding genome structure and function to analyse genotype-phenotype relationships
What are key genome analysis techniques?
Karyotyping
Genetic mapping
Genome sequenicng
What are common genome engineering methods?
Gene knockouts
CRISPR/Cas9 genome editing
What is genetic engineering?
Using recombinant DNA (rDNA) technology to alter an organism’s genetic makeup
What is recombinant DNA technology?
Comining DNA molecules from different organisms and inserting them into a host
What are the key steps in recombinant DNA techniques?
1 - Isolate gene of interest via PCR
2 - Cut DNA and plasmid with restriction enzymes
3 - Ligate GOI into plasmid vector
4 - Transform itno bacteria and select for growth
How does PCR isolate a Gene Of Interest?
Uses priemrs to amplify DNA exponentially
What do restriction enzymes do?
Cleave DNA at specific palindromic sequences
What is the difference between sticky and blunt ends?
Sticky: Overhangs ensure unidirectional insertion
Blunt: Both orientations possible
What is a plasmid vector?
A circular DNA molecule used to propagate inserted sequences
How is DNA cloned into a vector?
Both fragment and vector are cut with the same restriction enzyme ligated
What is blue/white seclection?
White colonies: Successful gene insertion (lacZ gene disrupted)
Blue colonies: no insert
What is the best way to study gene function?
Gene knockouts - removing a gene to assess phenotype
What is forward genetics?
Identifying mutations and linking them to a genotype
What is reverse genetics?
Using genome sequencing to determine gene function
What is transgenesis?
Introducing recombinant DNA into organisms
What are key methods for stable transgenesis?
Injecting DNA into the germline for inheritance
How does homologous recombination work?
Replacing endogenous genes with transfected sequences via recombination
How can transgenic techniques be used?
Functional genomics - assigning function to genes
Humanised models - replaing mouse/fly genes with human orthologs
GMO crops and livestock - enhanced yield, disease resistance
Medicine - insulin production, xenotransplantation
Why is CRISPR/Cas9 revolutionary?
Universal across species, accelerating genome engineering
What are ethical concerns in genetic engineering?
Designer babies
Unknown long-term effects of GMOs
Monoculture risks
