Thus, in the context of a particular type of variant, we raise awareness of the limitations of minigene splicing assays and emphasize the importance of sequence context in regulating splicing. In the pSP元 context, only 18 of the 20 wild-type minigene constructs generated the expected wild-type transcripts whereas 8 of the 18 (44%) corresponding variant minigene constructs generated wild-type transcripts. In the pET01 context, all 20 wild-type minigene constructs generated the expected wild-type transcripts of the 20 corresponding variant minigene constructs, 14 (70%) generated wild-type transcripts. We found a high discordance rate in terms of the generation of wild-type transcripts, not only between FLGSA and the minigene assays but also between the different minigene assays. Herein, we analyzed the splicing outcomes of 20 +2T>C variants that generate some wild-type transcripts in two minigene assays. 6Institute of Medical Genetics, School of Medicine, Cardiff University, Cardiff, United KingdomĬombining data derived from a meta-analysis of human disease-associated 5′ splice site GT>GC (i.e., +2T>C) variants and a cell culture-based full-length gene splicing assay (FLGSA) of forward engineered +2T>C substitutions, we recently estimated that ∼15–18% of +2T>C variants can generate up to 84% wild-type transcripts relative to their wild-type counterparts.5Laboratory of Excellence GR-Ex, Paris, France.4Service de Génétique Médicale et de Biologie de la Reproduction, CHRU Brest, Brest, France.3Univ Brest, Inserm, EFS, UMR 1078, GGB, Brest, France.2Shanghai Institute of Pancreatic Diseases, Shanghai, China.1Department of Gastroenterology, Changhai Hospital, Second Military Medical University, Shanghai, China.Cooper 6, Claude Férec 3,4, Zhuan Liao 1,2* and Jian-Min Chen 3* Jin-Huan Lin 1,2†, Hao Wu 1,2†, Wen-Bin Zou 1,2†, Emmanuelle Masson 3,4, Yann Fichou 3,5, Gerald Le Gac 3,4,5, David N.
0 Comments
Leave a Reply. |
AuthorWrite something about yourself. No need to be fancy, just an overview. ArchivesCategories |