bioRxivBioengineering
Decoupling tRNA promoter and processing activities enables specific Pol-II Cas9 guide RNA expression
D. J. Knapp, Y. S. Michaels, M. Jamilly, Q. R. Ferry, H. Barbosa, T. A. Milne, T. A. Fulga
Abstract
Spatial/temporal control of Cas9 guide RNA expression could considerably expand the utility of CRISPR-based technologies. Current approaches based on tRNA processing offer a promising strategy but suffer from high background. Here we developed a variant screening platform to identify differential sequence determinants of human tRNA promoter and processing activities. Rational design based on the ensuing principles allowed us to engineer an improved tRNA scaffold that enabled highly specific guide RNA production from a Pol-II promoter.
- organism: human
- assay: CRISPR screen
- research focus: gene regulatory network inference
- approach: experimental assay
Reproducibility ledger
Nobody has reported trying to reproduce this paper yet. Did you replicate an experiment or run the code? Your report saves the next lab a week.
Sign in to file a reproducibility report.
Related papers
- License to cut: Smart RNA guides for conditional control of CRISPR-Cas9
A. Baccouche, A. Adel, … A. J. Genot·bioRxiv·Oct 26, 2022
- Engineered transcription-associated Cas9 targeting in eukaryotic cells
Gregory W. Goldberg, Manjunatha Kogenaru, … Jef D. Boeke·Nature CommunicationsPeer-reviewed·Nov 27, 2024
- Specific Modulation of CRISPR Transcriptional Activators through RNA-Sensing Guide RNAs in Mammalian Cells and Zebrafish Embryos
Oana Pelea, Sarah Mayes, … Tatjana Sauka‐Spengler·eLifePeer-reviewed·Jul 8, 2025
- Multilayered regulation by RNA thermometers enables precise control of Cas9 expression
Elise K. Phillips, Dawn M. Klingeman, … William G. Alexander·bioRxiv·May 12, 2025
- Selection of Extended CRISPR RNAs with Enhanced Targeting and Specificity
A. Herring-Nicholas, H. Dimig, … E. A. Josephs·bioRxiv·Jan 11, 2023
- Controlling gene expression in mammalian cells using multiplexed conditional guide RNAs for Cas12a
Lukas Oesinghaus, Friedrich Christian Simmel·Angewandte Chemie International EditionPeer-reviewed·Sep 17, 2021