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Guide-free Cas9 is detrimental to pig preimplantation embryo development

ABSTRACT: Guide-free Cas9 is detrimental to pig preimplantation embryo development

PROVIDER: PRJNA930390 | ENA |

REPOSITORIES: ENA

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			Action	DRS
	SRR23302800_1.fastq.gz	Fastqsanger.gz
	SRR23302800_2.fastq.gz	Fastqsanger.gz
	SRR23302801_1.fastq.gz	Fastqsanger.gz
	SRR23302801_2.fastq.gz	Fastqsanger.gz
	SRR23302802_1.fastq.gz	Fastqsanger.gz

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Guide-free Cas9 is detrimental to pig preimplantation embryo development

Project description:Germline genome editing by the CRISPR/Cas9 system has potential for generation of genetically modified animals/the targeted correction of pathological mutations, understanding the mechanism of early embryonic development. Previous reports showed that Cas9 protein without sgRNAs could induce genomic damage in cell levels in vitro. However, whether the detrimental effects occurred in embryos after Cas9-treatment remains unknown. Here, by using pig embryos as the subjective, we found that Cas9 protein transcribed from injected Cas9 mRNA could persist to at least blastocyst stage. Cas9 protein alone could induce the genome damage of preimplantation embryos, represented by increased phosphorylated histone H2AX (pH2AX) foci on the chromatin fiber, leading to apoptosis and decreased cell number of blastocysts. In addition, single-blastocyst RNA sequencing confirmed that Cas9 protein without sgRNAs could also cause the blastocyst transcriptome changes, depressing the embryo development signal pathway, such as cell cycle, metabolism, and cellular communication related signal pathways, while activating apoptosis and necroptosis signal pathway, both of which together resulted in impaired the preimplantation embryonic development.

2023-02-05 | GSE224238 | GEO

Preimplantation mouse embryo development MOE430B

Project description:Preimplantation mouse embryo development on Affymetrix MOE430B. Zeng et al (2004) Dev Biol 272:483-496. Keywords: other

2004-09-10 | GSE1745 | GEO

Preimplantation mouse embryo development U74Av2

Project description:Preimplantation mouse embryo development study on Affymetrix U74Av2. Zeng et al Dev Biol (2004) 272:483-496. Keywords: other

2004-07-06 | GSE1391 | GEO

Preimplantation mouse embryo development MOE430A

Project description:Preimplantation mouse embryo development study on Affymetrix MOE430A. Zeng et al (2004) Dev Biol 272:483-496. Keywords: other

2004-09-10 | GSE1744 | GEO

Evaluation of guide-free Cas9-induced genomic damage and transcriptome changes in pig embryos.

Project description:Cas9 protein without sgRNAs can induce genomic damage at the cellular level in vitro. However, whether the detrimental effects occur in embryos after Cas9 treatment remains unknown. Here, using pig embryos as subjects, we observed that Cas9 protein transcribed from injected Cas9 mRNA can persist until at least the blastocyst stage. Cas9 protein alone can induce genome damage in preimplantation embryos, represented by the increased number of phosphorylated histone H2AX foci on the chromatin fiber, which led to apoptosis and decreased cell number of blastocysts. In addition, single-blastocyst RNA sequencing confirmed that Cas9 protein without sgRNAs can cause changes in the blastocyst transcriptome, depressing embryo development signal pathways, such as cell cycle, metabolism, and cellular communication-related signal pathways, while activating apoptosis and necroptosis signal pathways, which together resulted in impaired preimplantation embryonic development. These results indicated that attention should be given to the detrimental effects caused by the Cas9 protein when using CRISPR-Cas9 for germline genome editing, especially for the targeted correction of human pathological mutations using germline gene therapy.

| S-EPMC10551558 | biostudies-literature

Transcription profiling of preimplantation mouse embryo development

Project description:Preimplantation mouse embryo development study on Affymetrix MOE430A. Zeng et al (2004) Dev Biol 272:483-496.

2008-06-11 | E-GEOD-1744 | biostudies-arrayexpress

Transcriptomic analysis of mouse preimplantation embryo development and nutrient provision

Project description:Purpose: The goals of this study are to define transcriptome (RNA-seq) of mouse preimplantation embryos at different stages of development under a range of different environmental conditions. Methods: Mouse preimplantation embro transcriptional profiles were generated using embryos at several different developmental stages using Smart-seq2. Results: RNA-seq analysis finds that there is a highly dynamic pattern of gene expression during the preimplantation period. The sensitiivty to nutrient conditions varies markedly at different stages of development, with 2C embryos more sensitive to pyruvate omission than later stage embryos. Conclusions: Our study represents a comprehensive analysis of the mouse preimplantation development transcriptome, and how pyruvate provision impacts different developmental stages.

2020-10-14 | GSE159484 | GEO

Isoform-resolved transcriptome of the human preimplantation embryo [Illumina]

Project description:Human preimplantation development is a complex process involving extensive remodeling of gene expression. However, the preimplantation embryo transcriptome has only been annotated using short-read sequencing, which fails to capture full-length mRNAs and associated isoform diversity. We present a novel human embryo transcriptome using integrated long- and short-read RNA sequencing data. Our analysis reveals a total of 110,212 novel isoforms transcribed from known genes containing either a novel combination of known splice sites or at least one novel splice site, and 17,964 isoforms transcribed from completely novel genes located either in antisense direction of known genes or in intergenic space.

2023-09-26 | GSE190544 | GEO

Isoform-resolved transcriptome of the human preimplantation embryo [PacBio]

2023-09-26 | GSE190547 | GEO

High-resolution Ribosome Profiling Reveals Translational Selectivity in Bovine Preimplantation Embryo Development

Project description:High resolution polysome fractionation and low-input ribosome profiling of bovine oocytes and preimplantation embryos has enabled us to define the translational landscape of early embryo development at an unprecedented level. We systematically and comparatively analyzed the transcriptome, polysome- and nonpolysome-bound RNA profiles of bovine oocytes and early embryos at 2-, 8-cell, morula, and blastocyst stage, and defined four modes of translational selectivity in bovine preimplantation embryo development: i. selective translation of non-abundant mRNAs, ii. active translating highly expressed mRNAs, iii. Translationally suppressed abundant mRNAs, and iv. Monosomaly occupied mRNAs. A strong selection towards genes involved in mitochondrial function and metabolic pathways was found throughout bovine preimplantation development. We found translatome largely follows transcriptome at oocytes, followed by a marked translational control at 8-cell embryos, which is gradually synchronized at the morula and blastocyst stage. We identified important novel cellular/embryonic functional regulators that being utilized and prioritized for translation at each developmental stage, that accompanies little-known bovine embryonic developmental programming. Together, these data reveal a unique spatiotemporal translational regulation that accompanies bovine preimplantation development.

2022-10-01 | GSE196484 | GEO

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