Transcriptome sequencing (RNA-Seq) studies the complete set of all mRNAs transcribed by specific cells under a particular functional state. Next-generation high-throughput sequencing technology comprehensively and rapidly obtains almost all transcript sequence information from a specific tissue or organ of a species under a given condition. This enables accurate analysis of critical life science questions such as differential gene expression, gene structural variations, and screening of molecular markers (SNPs or SSRs).

1.Thirteen Years of Experience: Over a decade of expertise in transcriptome sequencing analysis. We have independently developed multiple biologically recognized algorithms, such as the alternative splicing detection algorithm ASD and CASH, achieving higher detection rates and accuracy than similar software.
2.Species-Agnostic Analysis: Not reliant on existing species information; capable of studying non-model organisms. We have established multiple pipelines tailored for data from different platforms.
3.Proprietary Cloud Platform: Novel Bio's self-developed transcriptome analysis cloud platform enables one-click data analysis, covering both basic and advanced analytical needs.
4.Integrated Databases & Proven Track Record: Integrated numerous academically recognized transcriptome-related databases, fundamentally enhancing the breadth and precision of downstream analysis. We have contributed to 300+ SCI publications, assisting research teams in publishing the world's first autistic monkey model in Nature.

Tissue Samples:
1.Animal tissue: >100 mg.
2.Plant tissue: >200 mg.
3.Cell sample: >5×10⁶ cells.
4.Whole blood: >2 mL.

RNA Samples:
1.Quantity required: >3 μg RNA.
2.Concentration: >50 ng/μL.
3.Purity:
·OD260/OD280 ratio between 1.8–2.2.
·OD260/OD230 ratio >2.
·28S/18S ratio >1.
·RIN >7.0 for animal samples; RIN >6.5 for plant samples.
·RNA must show no significant degradation.

Experimental Groups:
Transgenic (TG) cynomolgus monkeys (T14, T05, T07, T09) : Wild-type (WT) cynomolgus monkeys = 4 : 4
Primary Technical Methods:
Transcriptome sequencing, PCR, Southern blotting, Immunohistochemical staining, Western blotting, Metabolic mass spectrometry, etc.

Analysis Methods:
Differential expression gene screening analysis, Gene expression quantification analysis, Genomic integration site analysis, Behavioral data analysis, Copy number analysis, etc.

Autism is a disease caused by developmental disorders of the brain, characterized by difficulties in emotional expression, impaired social interaction, problems with verbal and non-verbal communication, and commonly exhibited restricted behaviors and repetitive movements, along with marked specific interests. Individuals often cannot perform normal language expression and social activities, frequently engaging in stereotyped and repetitive actions and behaviors. The cause of autism remains unknown, and many researchers suspect it is controlled by genes and triggered by environmental factors. Currently, there is no effective treatment for autism.

A major obstacle in autism research has been the lack of animal models that can faithfully recapitulate the core symptoms observed in human patients. While mouse models of autism exist, the complexity of the human brain and nervous system far surpasses that of rodents. Consequently, mouse models often fall short in studying human neurological and psychiatric disorders. To establish a more advanced animal model with a complex nervous system that better mimics the human condition, the research teams led by Qiu Zilong and Sun Qiang employed non-human primates - specifically, cynomolgus monkeys (Macaca fascicularis), whose neural systems share remarkable similarities with humans - to develop a model carrying the human MECP2 gene.

(A) Design scheme of the exogenous transgene fragment in the viral vector.
(B) Newborn F0 generation transgenic cynomolgus monkey under maternal care.
(C) Genome-wide mapping of the transgene insertion site obtained by deep sequencing.
(D) Schematic of movement path tracking results and statistics of repetitive path duration.
(E) Behavioral paradigm for testing social behavior within a group.
(F) Results of paired social behavior tests in the F0 generation.
(G) Newborn F1 generation transgenic cynomolgus monkey.
(H) Results of paired social behavior tests in the F1 generation.

his study first successfully constructed a transgenic monkey model carrying a human autism-related gene by introducing the human MECP2 gene into the genome of cynomolgus monkeys using lentiviral vector infection. The research team also successfully obtained a second generation of transgenic monkeys carrying the human MECP2 gene through testicular allograft transplantation. Behavioral studies revealed that these F1 generation monkeys also exhibited human autism-like characteristics, particularly in social behaviors, similar to the parental generation.

Through meticulous and rigorous molecular biology and genetic experiments, the research team demonstrated the successful integration and correct expression of the MECP2 transgene in the monkeys. Compelling and extensive animal behavioral experiments confirmed that the transgenic monkeys displayed significant human autism-like symptoms in stereotyped behaviors and social interactions.

This work can be considered a milestone achievement in the field of autism research, opening a new door for scientists to deeply investigate the mechanisms of autism and potentially conquer the disorder.

Novel Bio is honored to have participated in this outstanding research, contributing modestly to this study. To demonstrate the correct expression of the introduced MECP2 gene, researchers conducted comparative transcriptome analysis of brain cortex tissues from transgenic and wild-type monkeys. We performed RNA sequencing using the Life Technologies Ion Proton™ platform and conducted in-depth bioinformatics analysis. The results proved significant differences at the transcriptome level between the brain cortices of transgenic and wild-type monkeys, with genes related to metabolism and brain development showing markedly different expression patterns. This indicates that the introduced MECP2 gene was correctly expressed and regulated the expression of downstream genes.

Link to Original Article:oi:10.1038/nature16533