Browse
Submit Data
Databases
API
Help

Dataset Information

0 Views

0 Connections

0 Citations

0 Reanalyses

0 Downloads

Omics score: 0

ABSTRACT: Efficient in vivo prime editing corrects the most frequent phenylketonuria variant, associated with high unmet medical need

PROVIDER: PRJNA1015257 | ENA |

REPOSITORIES: ENA

ACCESS DATA

Json Xml

Dataset's files

Source:

			Action	DRS
	SRR26030800_1.fastq.gz	Fastqsanger.gz
	SRR26030800_2.fastq.gz	Fastqsanger.gz
	SRR26062800.fastq.gz	Fastqsanger.gz
	SRR26062900_1.fastq.gz	Fastqsanger.gz
	SRR26062900_2.fastq.gz	Fastqsanger.gz

Items per page:

1 - 5 of 1026

Similar Datasets

Treatment of a metabolic liver disease by in vivo prime editing in mice

Project description:Prime editing is a highly versatile CRISPR-based genome editing technology with the potential to correct the vast majority of genetic defects1. However, correction of a disease phenotype in vivo in somatic tissues has not been achieved yet. Here, we establish proof-of-concept for in vivo prime editing, that resulted in rescue of a metabolic liver disease. We first develop a size-reduced prime editor (PE) lacking the RNaseH domain of the reverse transcriptase (SpCas9-PERnH), and a linker- and NLS-optimized intein-split PE construct (SpCas9-PE p.1153) for delivery by adeno-associated viruses (AAV). Systemic dual AAV-mediated delivery of this variant in neonatal mice enables installation of a transversion mutation at the Dnmt1 locus with 15% efficiency on average. Next, we targeted the disease-causing mutation in the phenylalanine hydroxylase (Pah)enu2 mouse model for phenylketonuria (PKU). Correction rates of 1.5% using the dual AAV approach could be increased to up to 14% by delivery of full-length SpCas9-PE via adenoviral vector 5 (AdV5), leading to full restoration of physiological blood phenylalanine (L-Phe) levels below 120 µmol/L. Our study demonstrates in vivo prime editing in the liver at two independent loci, emphasizing the potential of PEs for future therapeutic applications.

2021-12-29 | GSE174757 | GEO

Project description:A base editing strategy using mRNA-LNPs for in vivo correction of the most frequent phenylketonuria variant

| PRJNA1026949 | ENA

Mutation-specific reporter for optimization and enrichment of prime editing

Project description:Prime editing is a versatile genome-editing technique that shows great promise for the generation and repair of patient mutations. However, some genomic sites are difficult to edit and optimal design of prime-editing tools remains elusive. Here we present a fluorescent prime editing and enrichment reporter (fluoPEER), which can be tailored to any genomic target site. This system rapidly and faithfully ranks the efficiency of prime edit guide RNAs (pegRNAs) combined with any prime editor variant. We apply fluoPEER to instruct correction of pathogenic variants in patient cells and find that plasmid-editing enriches for genomic editing up to 3-fold compared to conventional enrichment strategies. DNA repair and cell cycle-related genes are enriched in the transcriptome of edited cells. Stalling cells in the G1/S boundary increases prime editing efficiency up to 30%. Together, our results show that fluoPEER can be employed for rapid and efficient correction of patient cells, selection of gene-edited cells, and elucidation of cellular mechanisms needed for successful prime editing.

2022-02-03 | GSE195977 | GEO

Improved split prime editors enable efficient in vivo genome editing

Project description:Improved split prime editors enable efficient in vivo genome editing

| PRJNA1128556 | ENA

Prime editing for functional repair in patient-derived disease models

Project description:Prime editing is a novel genome editing technology using fusion proteins of Cas9-nickase and reverse transcriptase, that holds promise to correct a wide variety of genetic defects. We succeeded in efficient prime editing and functional recovery of disease-causing mutations in patient-derived liver and intestinal stem cell organoids. Whole genome sequencing of did not detect off-target mutations or a mutational signature induced by prime editing.

| EGAS00001004611 | EGA

Improved split prime editors enable efficient in vivo genome editing [Amplicon_Seq_in_vitro]

Project description:Improved split prime editors enable efficient in vivo genome editing [Amplicon_Seq_in_vitro]

| PRJNA1128634 | ENA

Project description:Engineering CjCas9 for Efficient Base Editing and Prime Editing

| PRJNA1056772 | ENA

Project description:Prime Editing Corrects RBM20-Mediated Dilated Cardiomyopathy in Human Cardiomyocytes

| PRJNA1285140 | ENA

Genotoxic effects of base and prime editing in human hematopoietic stem cells [NHEJ_PrimeE]

Project description:Emerging base and prime editing may provide safer and more precise genetic engineering than nuclease-based approaches bypassing the dependence on DNA double strand breaks (DSBs). However, little is known about cellular responses and genotoxicity. Here, we comparatively assessed state-of-the-art base and prime editors (B/PE) versus Cas9 in human hematopoietic stem/progenitor cells (HSPCs). BE and PE induced detrimental transcriptional responses constraining editing efficiency and/or HSPC repopulation in xenotransplants, albeit to a lesser extent than Cas9. DNA DSBs and their genotoxic byproducts, including deletions and translocations, were less frequent but not abrogated by BE and PE, particularly for cytidine BE due to suboptimal inhibition of base excision repair. Tailoring timing and B/PE expression enabled highly efficient and precise editing of long-term repopulating HSPCs. However, we uncovered a genome-wide effect of BEs on the mutational landscape of HSPCs, raising concerns for a potential genotoxic impact and calling for further investigations and improvements in view of clinical application.

2023-07-18 | GSE220765 | GEO

Genotoxic effects of base and prime editing in human hematopoietic stem cells [NHEJ_BaseE]

2023-07-18 | GSE220763 | GEO