1 2 3 4 5 6 7

The blockage of the Tyrosine Phosphatase Gamma, or Ptpg, receptor was performed using antisense technique. Strategies using antisense techniques to inhibit expression of specific genes have now been used for over a decade. Antisense technique is different from the successful method of antigene strategy, which is based on homologous recombination and could result in being very time consuming. Antisense oligonucleotide design provides synthesis and results, which are reasonably simple and fast.
Antisense Oligonucleotides (ONs) are short sequences of single stranded DNA, most likely less than 30 base pairs, which are engineered through chemical synthesis. The antisense ONs' sequence is specifically made to be complementary to an intracellular targeted mRNA molecule. The ONs have a high negative density charge; therefore the mechanism for the travel through the plasma membrane is still unclear. But, strategies have been developed to increase the potency of the antisense ONs within the cell. On such strategy is the use of a cationic liposome, which is found in the Lipofection used in this experiment. Two types of oligonucleotides were used in this experiment. The control embryos were placed in a solution containing an ON with a scrambled sequence of DNA and therefore was unable to bind to Ptp
g. The experimental embryos were exposed to the antisense ON for Ptpgand therefore the gene's expression was blocked.

©Cebra-Thomas, 2001

Last Modified: 1 May 2001

[Lab Protocols | Students | Cebra-Thomas | Course | Links ]