Reverse & forward Genetics

Question 1

Reverse genetics

Answer 1

from genotype to phenotype

Question 2

why reverse genetics

Answer 2

Understanding the phenotypic effects of specific gene sequences

Question 3

Reverse genetic Method

Answer 3

1) Identify gene of interest
2) mutate it
3) characterize effects

Question 4

How do you mutate a gene of interest (Reverse Genetics)

Answer 4

1) gene knockout, disruption, knockdown with CRISPR-CAS
2) Reducing expression using interference RNA (RNAi)
3) Overexpression (disruption supression genes or silencers)

Question 5

Advantages for reverse genetics

Answer 5

1) more control
2) can be used to validate findings
3) can be used to generate animal models
4) studying disease mechanisms

Question 6

Limitations of Reverse genetics

Answer 6

1) off target effects
2)incomplete phenotyic penetrance (it does not show even though it is mutated)
3) gene-environment interaction can complicate matters
4) limited understanding of gene functions
5) resource intensive

Question 7

Forward genetics

Answer 7

From phenotype to genotype

Question 8

Why use forward genetics

Answer 8

When you do not know what is causing the disease

Question 9

method Forward genetics

Answer 9

1) identify phenotype of interest
2) find an appropriate model system
3) discriminate between wildtype
4) introduce random mutations
5) target genes
6) screen for mutations
7) check if it is heritable
8) identify mutated gene
9) sequencing
10) mapping

Question 10

Advantages forward genetics

Answer 10

+ useful for the discovery of novel genes
+ identifying casual variants
+ can lead to insights in biological mechanisms
+ can be relevant for human disease

Question 11

Limitations forward genetics

Answer 11

• Labor and resource intensive
• Genetic background can make this more complicated and lead to confounders
• Limited resolution