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Macaca mulatta

ABSTRACT: TALEN-mediated MeCP2 Gene Mutagenesis in Rhesus and Cynomolgus Monkeys

PROVIDER: PRJNA230664 | ENA |

REPOSITORIES: ENA

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			Action	DRS
	SRR1046130.fastq.gz	Fastqsanger.gz
	SRR1047650.fastq.gz	Fastqsanger.gz
	SRR1047660.fastq.gz	Fastqsanger.gz
	SRR1047641.fastq.gz	Fastqsanger.gz
	SRR1047651.fastq.gz	Fastqsanger.gz

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TALEN-mediated MeCP2 Gene Mutagenesis in Rhesus and Cynomolgus Monkeys

Project description:We report Transcription Activator-like Effector Nucleases (TALEN)-mediated mutagenesis of the Rett Syndrome gene, methyl-CpG binding protein 2 (Mecp2), in nonhuman primates (NHPs). Microinjection of Mecp2-targeting TALEN plasmids into rhesus and cynomolgus zygotes caused 40-50% of the embryos harboring mutations when examined beyond 8-cell stage. The surrogate pregnancy rates were 28.6% (2/7) and 16.7% (1/6) for rhesus and cynomolgus, respectively. The 2 rhesus surrogates went through miscarriage at 30- and 63/64-days of gestation. Analyses of two 63/64-day-old liter-mate embryos indicated both were male and carried MeCP2 mutations, yet chimeric. Mutation rate was higher in one embryonic brain than the other. RNA sequencing and anatomical analyses indicated that the brain with higher-degree mutations was underdeveloped, which, together with mid-gestation-stage male lethality likely recapitulate the disease. One live delivery of a female cynomolgus monkey occurred after 162 days of gestation, with abundant mutagenesis in peripheral tissues examined. We conclude that TALEN-mediated mutagenesis greatly facilitates disease modeling in NHPs.

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Project description:Splenic tissue was isolated from four adult male Indian-origin Rhesus monkeys serologically positive for non-pathogenic SHIV 89.6 and from matched uninfected four adult male Indian-origin Rhesus monkeys respectively. The corresponding RNA was processed by cDNA microarray analysis. Sample RNA extracted from the tissue samples. Reference RNA was prepared from a pool of Rhesus monkeys' RNA. Microarray hybridization was carried out by labeling Reference RNA as Cy3 and sample RNA as Cy5. The sample was incubated at 55C for 16hr.

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