Regional random mutagenesis driven by multiple sgRNAs and diverse on-target genome editing events to identify functionally important elements in non-coding regions DOI Creative Commons
Kento Morimoto, Hayate Suzuki, Akihiro Kuno

et al.

Open Biology, Journal Year: 2024, Volume and Issue: 14(4)

Published: April 1, 2024

Functional regions that regulate biological phenomena are interspersed throughout eukaryotic genomes. The most definitive approach for identifying such is to confirm the phenotype of cells or organisms in which specific have been mutated removed from genome. This invaluable functional analysis genes with a defined element, protein-coding sequence. By contrast, no platforms established study cis -elements microRNA cluster consisting multiple microRNAs overlap. Whole-genome mutagenesis approaches, as via N -ethyl- -nitrosourea and gene trapping, greatly contributed elucidating function coding genes. These methods almost never induce deletions genomic mutations within narrow region. In other words, clusters cannot be effectively targeted manner. Herein, we novel region-specific random method named CRISPR- transposase-based regional (CTRL-mutagenesis). We demonstrate CTRL-mutagenesis randomly induces diverse target murine embryonic stem cells. Comparative mutants harbouring subtly different same region would facilitate further -element clusters.

Language: Английский

Regional random mutagenesis driven by multiple sgRNAs and diverse on-target genome editing events to identify functionally important elements in non-coding regions DOI Creative Commons
Kento Morimoto, Hayate Suzuki, Akihiro Kuno

et al.

Open Biology, Journal Year: 2024, Volume and Issue: 14(4)

Published: April 1, 2024

Functional regions that regulate biological phenomena are interspersed throughout eukaryotic genomes. The most definitive approach for identifying such is to confirm the phenotype of cells or organisms in which specific have been mutated removed from genome. This invaluable functional analysis genes with a defined element, protein-coding sequence. By contrast, no platforms established study cis -elements microRNA cluster consisting multiple microRNAs overlap. Whole-genome mutagenesis approaches, as via N -ethyl- -nitrosourea and gene trapping, greatly contributed elucidating function coding genes. These methods almost never induce deletions genomic mutations within narrow region. In other words, clusters cannot be effectively targeted manner. Herein, we novel region-specific random method named CRISPR- transposase-based regional (CTRL-mutagenesis). We demonstrate CTRL-mutagenesis randomly induces diverse target murine embryonic stem cells. Comparative mutants harbouring subtly different same region would facilitate further -element clusters.

Language: Английский

Citations

1