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An antibody independent verification method quantifies m6A in the chicken β-actin zipcode

ABSTRACT: An antibody independent verification method quantifies m6A in the chicken β-actin zipcode

PROVIDER: PRJNA767616 | ENA |

REPOSITORIES: ENA

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			Action	DRS
	SRR16127985_1.fastq.gz	Fastqsanger.gz
	SRR16127985_2.fastq.gz	Fastqsanger.gz
	SRR16127986_1.fastq.gz	Fastqsanger.gz
	SRR16127986_2.fastq.gz	Fastqsanger.gz
	SRR16127987_1.fastq.gz	Fastqsanger.gz

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An antibody independent verification method quantifies m6A in the chicken β-actin zipcode

Project description:N6-Methyladenosine (m6A) in mRNA regulates almost every stage in the mRNA life cycle, and the development of the high throughput detection of methylated sites in mRNA using MeRIPSeq or miCLIP revolutionized the m6A research field. Both methods are based on immunoprecipitation of fragmented mRNA. However, it is well documented that antibodies often have nonspecific activities, thus verification of identified m6A sites using an antibody-independent method would be highly desirable. Currently such approaches are limited. Here we present RedBaron, an improved biochemical method for the site-specific detection and quantification of m6A in RNA. We demonstrate that the RedBaron method is able to accurately quantify m6A levels at specific transcripts in vivo. We used this assay for the site-specific detection and quantification of m6A within the chicken β-actin (ACTB) zipcode sequence in chicken embryos and in fibroblast cells. We demonstrate that methylation of this site in the β-actin zipcode enhances ZBP1 binding in vitro, whilst methylation of a nearby adenosine abolishes

2023-05-24 | GSE185078 | GEO

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Actin binding proteins expression levels when modifying the actin sequence

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RNA-Seq analysis of beta-actin knockout MEFs expressing NLS-tagged beta-actin

Project description:RNA-Seq analysis of beta-actin knockout MEFs expressing NLS-tagged beta-actin to study role of actin in transcription

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Replacing skeletal muscle alpha-actin with cardiac actin in mouse skeletal muscle

Project description:Skeletal muscle actin mice (Crawford et al., (2002) Mol Cell Biol 22, 5587) were crossed with cardiac actin transgenic mice (termed "ACTC^Coco" or "Coco" for short), to produce mice that had cardiac actin instead of skeletal muscle actin in their skeletal muscles (termed "ACTC^Co/KO" or for short "Coco/KO"). Microarray analysis using the Illumina mouse-6 v1.1 expression beadchip was performed on RNA extraced from the soleus muscle of Coco/KO mice and wildtype mice, to confirm the swith in actin isoform expression, and to determine what other differences might exist between wildtype mice and the Coco/KO mice. Keywords: genetic modification 3 RNA samples (each being the pool of two individual samples extracted from different soleus muscles from different individual mice) per genotype (either wildtype or Coco/KO) were used. The total 6 RNA samples were processed using an Illumina mouse-6 v1.1expression beadchip and then the differentially expressed genes determined.

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