Autism-like behaviours and germline transmission in transgenic monkeys overexpressing MeCP2 

   Transcriptome analysis for transgenic monkey. Total RNA was extracted from three independent pieces of cortical tissues from brains of T05, T07, T09 and T14 and four WT monkeys by Trizol reagent (Invitrogen) separately. The RNA quality was checked by Bioanalyzer 2200 (Aligent) and kept at −80 °C. The RNA with RIN(RNA integrity number) > 8.0 is acceptable for cDNA library construction. RNAseq and bioinformatic data analysis were performed by Shanghai Novelbio Ltd.

    Library construction and RNA-seq. The cDNA libraries for single-end sequencing were prepared using Ion Total RNA-Seq Kit v2.0 (Life Technologies) according to the manufacturer’s instructions. The cDNA libraries were then processed for the proton sequencing process according to the commercially available protocols. Samples were diluted and mixed, the mixture was processed on a OneTouch 2 instrument (Life Technologies) and enriched on a OneTouch 2 ES station (Life Technologies) for preparing the template-positive Ion PI Ion Sphere Particles(Life Technologies) according to Ion PI Template OT2 200 Kit v2.0 (Life Technologies). After enrichment, the mixed template-positive Ion PI Ion Sphere Particles of samples was loaded on to 1 P1v2 Proton Chip (Life Technologies) and sequenced on Proton Sequencers according to Ion PI Sequencing 200 Kit v2.0 (Life Technologies).

    Acknowledgements We thank M.-m. Poo for comments on the manuscript,Y.-Z. Li, Y.-Z. Lu, F. Liu and X. Zhou for maintaining monkey colony, W. Lu,X.-H. Guo and Y. F. Zhou of Fudan Children’s Hospital for assistance in mass spectrometry and electroencephalogram, D. Chen and B. Zhang of Novel Bioinformatics for RNA-seq data analysis, and C.-H. Li of Shanghai Geneskies Company for analysis of genomic integration sites.