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High Level Expression of Transgenesis in Animal Cells - High level expression of transgenes in cultured mammalian cells is used for the production of therapeutic recombinant proteins. Expression in mammalian cells is particularly desirable when characteristic mammalian glycosylation of the proteins is necessary for their therapeutic application.

The following factors contribute to a high-level expression.

1. Use of a strong enhancer/promoter, e.g., SV40 enhancer/early promoter, etc.

2. Lack of secondary structure within the 5' -untranslated region of the transgene mRNA.

3. Lack of an initiation codon in the 5'-untranslated region, i.e., before the correct initiation codon.

In view of the items 2 and 3, the 5'-untranslated region of mRNA should be as short as possible.

4. The sequence around the initiation codon should conform to Kozak's rules. Of these rules, the most important is the presence of a purin, at the -3 position and a G at the +4 position. The A of the AUG initiation codon is counted as +1, U as +2 and so on.

5. Absence of AU-rich sequences in the 3'-untranslated region of mRNA since this sequence reduces mRNA stability.

 
 

 
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