CRISPR-based dissection of miRNA binding sites using isogenic cell lines is hampered by pervasive noise.

ABSTRACT: Non-coding regulatory sequences play essential roles in adjusting gene output to cellular needs and are thus critical to animal development and health. Numerous such sequences have been identified in mammalian genomes ranging from transcription factors binding motifs to recognition sites for RNA-binding proteins and non-coding RNAs. The advent of CRISPR has raised the possibility of assigning functionality to individual endogenous regulatory sites by facilitating the generation of isogenic cell lines that differ by a defined set of genetic modifications. Here we investigate the usefulness of this approach to assign function to individual miRNA binding sites. We find that the process of generating isogenic pairs of mammalian cell lines with CRISPR-mediated mutations introduces extensive mol

SUBMITTER: Prajapat MK 

PROVIDER: S-EPMC11398363 | biostudies-literature | 2024 Sep

REPOSITORIES: biostudies-literature