Introduction to Recombinant Genetics- Biology 350

Inhibition of gene expression

Knockout organisms

Tutorial on Gene knockout mice.

Question:
If the organism, in which you are going to knock the gene out, has a ploidy level greater than haploid, how easy would it be to obtain a complete knockout, where there is no gene expressed? How would you do that?

Antisense RNA

Tutorial on Antisense RNA inhibition.

Inject complementary RNA or transfect with a construct that will express antisense RNA.

Flavr-Savr Tomato

Polygalacturonase (PG) causes softening of tomato as pectin is broken down during ripening.

Use of complementary PNA to inhibit gene expression. (Wikipedia)

Question:
As above, if the organism, in which you are going to knock the gene out, has a high ploidy level, how easy would it be to completely inhibit gene expressed using antisense RNA?

Rna interference

Nature reviews - RNA Interference tutorial and animation.

Question:
How much of a gene sequence would you need to know in order to develop an RNAi strategy? Are there potential pitfalls?