Split dCas12a activator for lncRNA H19 activation to enhance BMSC differentiation and promote calvarial bone healing

Nuong Thi Kieu Nguyen, Yi Tu, Hsiang Sheng Lee, Vu Anh Truong, Yi Hao Chang, Nam Ngoc Pham, Chin Wei Chang, Ya Hui Lin, Po Liang Lai, Pin Hsin Chen, Yelena V. Parfyonova, Mikhail Menshikov, Yu Han Chang*, Yu Chen Hu*

*Corresponding author for this work

Research output: Contribution to journalJournal Article peer-review

2 Scopus citations

Abstract

Healing of large calvarial bone defects in adults is challenging. We previously showed that inducing chondrogenic differentiation of mesenchymal stem cells from bone marrow (BMSC) or adipose tissue (ASC) before implantation can switch the repair pathway and improve calvarial bone healing. Split dCas12a activator is a new CRISPR activation system comprising the amino (N) and carboxyl (C) fragments of dCas12a protein, each being fused with synthetic transcription activators at both termini. The split dCas12a activator was shown to induce programmable gene expression in cell lines. Here we exploited the split dCas12a activator to activate the expression of chondroinductive long non-coding RNA H19. We showed that co-expression of the split N- and C-fragments resulted in spontaneous dimerization, which elicited stronger activation of H19 than full-length dCas12a activator in rat BMSC and ASC. We further packaged the entire split dCas12a activator system (13.2 kb) into a hybrid baculovirus vector, which enhanced and prolonged H19 activation for at least 14 days in BMSC and ASC. The extended H19 activation elicited potent chondrogenic differentiation and inhibited adipogenesis. Consequently, the engineered BMSC promoted in vitro cartilage formation and augmented calvarial bone healing in rats. These data implicated the potentials of the split dCas12a activator for stem cell engineering and regenerative medicine.

Original languageEnglish
Article number122106
Pages (from-to)122106
JournalBiomaterials
Volume297
DOIs
StatePublished - 06 2023
Externally publishedYes

Bibliographical note

Copyright © 2023 Elsevier Ltd. All rights reserved.

Keywords

  • Bone healing
  • CRISPRa
  • Cre/loxP
  • Gene delivery
  • Long non-coding RNA
  • Split dCas12a activator
  • Rats
  • Mesenchymal Stem Cells/metabolism
  • RNA, Long Noncoding/genetics
  • Animals
  • Cell Differentiation/genetics
  • Osteogenesis/genetics
  • Adipose Tissue

Fingerprint

Dive into the research topics of 'Split dCas12a activator for lncRNA H19 activation to enhance BMSC differentiation and promote calvarial bone healing'. Together they form a unique fingerprint.

Cite this