Biomolecular condensation of human IDRs initiates endogenous transcription via intrachromosomal looping or high-density promoter localization

Nucleic Acids Research, Год журнала: 2025, Номер 53(4)

Опубликована: Янв. 22, 2025

Protein intrinsically disordered regions (IDRs) are critical gene-regulatory components and aberrant fusions between IDRs DNA-binding/chromatin-associating domains cause diverse human cancers. Despite this importance, how influence gene expression, IDR fusion proteins provoke oncogenesis, remains incompletely understood. Here we develop a series of synthetic dCas9-IDR to establish that locus-specific recruitment can be sufficient stimulate endogenous expression. Using dCas9 fused the paradigmatic leukemogenic NUP98 IDR, also demonstrate activate transcription via localized biomolecular condensation in manner is dependent upon overall concentration, local binding density, amino acid composition. To better clarify oncogenic role IDRs, construct clinically observed show that, while generally non-overlapping, NUP98-IDR convergently drive core expression program donor-derived hematopoietic stem cells. Interestingly, find leukemic arises through differing mechanistic routes based partner; either distributed intragenic intrachromosomal looping, or dense at promoters. Altogether, our studies roles and, for connect capacity pathological cellular programs, creating potential opportunities generalized mechanistically tailored therapies.

Язык: Английский

Compact transcription factor cassettes generate functional, engraftable motor neurons by direct conversion

Cell Systems, Год журнала: 2025, Номер unknown, С. 101206 - 101206

Опубликована: Март 1, 2025

Язык: Английский

High-resolution profiling reveals coupled transcriptional and translational regulation of transgenes

bioRxiv (Cold Spring Harbor Laboratory), Год журнала: 2024, Номер unknown

Опубликована: Ноя. 26, 2024

Abstract Concentrations of RNAs and proteins provide important determinants cell fate. Robust gene circuit design requires an understanding how the combined actions individual genetic components influence both mRNA protein levels. Here, we simultaneously measure levels in single cells using HCR Flow-FISH for a set commonly used synthetic promoters. We find that promoters generate differences abundance effective translation rate these transcripts. Stronger not only transcribe more RNA but also show higher rates. While strength promoter is largely preserved upon genome integration with identical elements, choice polyadenylation signal coding sequence can large profiles mRNAs proteins. long-read direct sequencing to characterize full-length isoforms observe remarkable uniformity from transgenic units. Together, our high-resolution profiling offers insight into impact common on transcriptional translational mechanisms. By developing novel framework quantifying expression transgenes, have established system comparing native regulation building robust systems.

Язык: Английский