Gene function Flashcards
Homology
The similarity between 2 objects, due to a common ancestry
orthologous genes
Genes present in two different species, arising from a common ancestor that underwent speciation
Paralogous genes
Genes present in the same organism, arising from the duplication of an ancestral gene
sequence alignment
Looking in databases for known genes sharing substantial similarity
Both nucleotide and amino acid sequences
More sophisticated for amino acids
approaches to inactivate a gene
Homologous recombination
RNA interference
CRISPR
Homologous recombination (for gene inactivation)
- Endogenous portion of DNA is replaced by exogenous DNA segment
- The exogenous DNA (deletion cassette) is comprised between 2 homology arms. Having the same sequence of the DNA flanking the gene that has to be deleted
- The deletion cassette usually encodes for a selectable marker gene (bijv. antibiotic resistance)
- The exogenous gene will be expressed, allowing selection of knockout cells
RNA interference
Silences a gene, in stead of disturbing it.
- Based on the use of short RNA molecules that are complementary to the target RNA
- they are produced starting from a longer double-stranded RNA procursor
- These short RNAs (siRNAs) are loaded inside a protein called argonaute (Ago) that is part of the RNA-induced Silencing Complex (RISC)
- Ago, guided by the siRNA, introduces a cut in the target RNA, inducing its degradation
Gene knockdown
The number of RNA molecules for the gene of interest will be drastically reduced, but some residual RNA will still be present