CRISPR-dCas13-tracing reveals transcriptional memory and limited mRNA export in developing zebrafish embryos.

ABSTRACT:

Background

Understanding gene transcription and mRNA-protein (mRNP) dynamics in single cells in a multicellular organism has been challenging. The catalytically dead CRISPR-Cas13 (dCas13) system has been used to visualize RNAs in live cells without genetic manipulation. We optimize this system to track developmentally expressed mRNAs in zebrafish embryos and to understand features of endogenous transcription kinetics and mRNP export.

Results

We report that zygotic microinjection of purified CRISPR-dCas13-fluorescent proteins and modified guide RNAs allows single- and dual-color tracking of developmentally expressed mRNAs in zebrafish embryos from zygotic genome activation (ZGA) until early segmentation period without genetic manipulation. Using this approach, we uncover non-

SUBMITTER: Huang Y 

PROVIDER: S-EPMC9854193 | biostudies-literature | 2023 Jan

REPOSITORIES: biostudies-literature