Division 1.2.2.3 Information on the expression of the insert(s)

The applicant shall provide information:

• to demonstrate whether the inserted/modified sequence results in intended changes at the protein, RNA and/or metabolite levels;

• to characterise the potential unintended expression of new ORFs identified under point 1.2.2.2(f) as raising a safety concern.

For those purposes, the applicant shall provide the following information:

1. (a)

   The method(s) used for expression analysis together with their performance characteristics;

2. (b)

   Information on developmental expression of the insert during the life cycle of the plant.

   The requirement for information on developmental expression shall be considered on a case-by-case basis taking into account the promoter used, the intended effect(s) of the modification and scope of the application;

3. (c)

   Parts of the plant where the insert/modified sequences are expressed;

4. (d)

   Potential unintended expression of new ORFs identified under point 1.2.2.2(f) which raise a safety concern;

5. (e)

   Protein expression data, including the raw data, obtained from field trials and related to the conditions in which the crop is grown.

   Data on expression levels from those parts of the plant used for food and feed purposes shall be provided in all cases.

   In addition, information shall also be provided on the expression of target genes in other parts of the plant when tissue-specific promoters have been used and when this is relevant for the safety assessment. The minimum requirement for protein expression shall be data provided from three growing sites or from one site over three seasons. Permutations of the sites and seasons shall be acceptable provided that the minimum requirement is met. When justified by the nature of the insert (such as silencing approaches or where biochemical pathways have been intentionally modified), specific RNA(s) or metabolite(s) shall be analysed.

   For silencing approaches by RNAi expression, potential ‘off target’ genes should be searched by in silico analysis to assess if the genetic modification could affect the expression of other genes which raise safety concerns;

6. (f)

   With regard to the stacking of transformation events by conventional crossing, expression data shall be provided to assess the potential interactions between the events which may raise any additional safety concerns over protein and trait expression compared with the single transformation events. The comparison shall be carried out with data obtained from plants grown in the same field trials. On a case-by-case basis, and where concerns arise, additional information may be necessary.