Tunable, self-contained gene dosage control via proteolytic cleavage of CRISPR-Cas systems DOI Creative Commons
Noa Katz,

Connie An,

Yu‐Ju Lee

et al.

bioRxiv (Cold Spring Harbor Laboratory), Journal Year: 2024, Volume and Issue: unknown

Published: Oct. 9, 2024

Gene therapy holds great therapeutic potential. Yet, controlling cargo expression in single cells is limited due to the variability of delivery methods. We implement an incoherent feedforward loop based on proteolytic cleavage CRISPR-Cas activation or inhibition systems reduce gene against vector delivery. demonstrate dosage control for and inhibition, post-delivery tuning, RNA-based delivery, a genome-integrated marker. then target

Language: Английский

Compact transcription factor cassettes generate functional, engraftable motor neurons by direct conversion DOI
Nathan Wang, Honour O Adewumi, Brittany A Lende-Dorn

et al.

Cell Systems, Journal Year: 2025, Volume and Issue: unknown, P. 101206 - 101206

Published: March 1, 2025

Language: Английский

Citations

1
High-resolution profiling reveals coupled transcriptional and translational regulation of transgenes DOI Creative Commons
Emma L. Peterman, Deon Ploessl, Kasey S. Love

et al.

bioRxiv (Cold Spring Harbor Laboratory), Journal Year: 2024, Volume and Issue: unknown

Published: Nov. 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.

Language: Английский

Citations

2
Tunable, self-contained gene dosage control via proteolytic cleavage of CRISPR-Cas systems DOI Creative Commons
Noa Katz,

Connie An,

Yu‐Ju Lee

et al.

bioRxiv (Cold Spring Harbor Laboratory), Journal Year: 2024, Volume and Issue: unknown

Published: Oct. 9, 2024

Gene therapy holds great therapeutic potential. Yet, controlling cargo expression in single cells is limited due to the variability of delivery methods. We implement an incoherent feedforward loop based on proteolytic cleavage CRISPR-Cas activation or inhibition systems reduce gene against vector delivery. demonstrate dosage control for and inhibition, post-delivery tuning, RNA-based delivery, a genome-integrated marker. then target

Language: Английский

Citations

0