We have a new article in the STAR Protocols journal

We’re excited to present our newest paper, Protocol for editing fibroblasts with in vitro transcribed Cas9 mRNA and profile off-target editing by optimized GUIDE-seq, published in the STAR Protocols journal in December 2023.

Highlights and Summary

  • A protocol for in vitro transcription of Cas9 mRNA with quality control step
  • Electroporation transfection to edit fibroblast with Cas9 mRNA
  • Optimization of dsODN concentration used for GUIDE-seq
  • Profile off-target editing by optimized GUIDE-seq method

CRISPR-Cas9 gene editing is an efficient technique to modify specific sites/regions of DNA. Delivery of the Cas9 by mRNA is particularly promising in pre-clinical genome editing applications for its transient, nonintegrating feature.

However, the off-target of Cas9-gRNA still remains a concern and needs a specific monitor. Here, we present a revised protocol to edit fibroblasts by in vitro transcribed Cas9 mRNA and profile its off-target effect by the optimized GUIDE-seq method. This protocol can also be applied to other cell lines.

Graphical Abstract
Graphical Abstract

Latest news

The event took place at the beautiful Oscarborg Fortress.
We have a post-doc position currently open, focusing on single-cell sequencing of the CRISPR-corrected cells.
Kata went to American Society of Gene and Cell Therapy (ASGCT) conference in Baltimore, which focuses on novel translational findings in the field.
Shiva presented her poster at the 2024 CRISPR Medicine Conference in Denmark.
We had the pleasure of attending the 41st NSI Annual Meeting on November 24th, 2023!