Robust genome and RNA editing via CRISPR nucleases in PiggyBac systems

Yuqian Jiang, Rachel Catherine Hoenisch, Yun Chang, Xiaoping Bao, Craig E. Cameron, Xiaojun Lance Lian

Research output: Contribution to journalArticlepeer-review

5 Scopus citations


CRISPR/Cas-mediated genome editing in human pluripotent stem cells (hPSCs) offers unprecedented opportunities for developing in vitro disease modeling, drug screening and cell-based therapies. To efficiently deliver the CRISPR components, here we developed two all-in-one vectors containing Cas9/gRNA and inducible Cas13d/gRNA cassettes for robust genome editing and RNA interference respectively. These vectors utilized the PiggyBac transposon system, which allows stable expression of CRISPR components in hPSCs. The Cas9 vector PB-CRISPR exhibited high efficiency (up to 99%) of inducing gene knockout in both protein-coding genes and long non-coding RNAs. The other inducible Cas13d vector achieved extremely high efficiency in RNA knockdown (98% knockdown for CD90) with optimized gRNA designs. Taken together, our PiggyBac CRISPR vectors can serve as powerful toolkits for studying gene functions in hPSCs.

Original languageEnglish (US)
Pages (from-to)313-320
Number of pages8
JournalBioactive Materials
StatePublished - Aug 2022

All Science Journal Classification (ASJC) codes

  • Biotechnology
  • Biomaterials
  • Biomedical Engineering


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