Human pluripotent stem cells (hPSCs) are a vital tool not only in research and disease modeling but also in developmental biology and regenerative medicine because of their ability to self-renew and differentiate. Current genome editing tactics in hPSCs require the incorporation of deleterious double-stranded DNA breaks followed by inefficient homology-directed repair (HDR). These methods can result in DNA insertion or deletion issues, apoptosis, oncogenic mutations and other negative outcomes.
Researchers at Arizona State University have developed a novel method using CRISPR tools to co-target genomic loci and an episomal reporter to enable single-nucleotide genomic changes in hPSCs without HDR. This method, called base-edited isogenic hPSC line generation using a transient reporter for editing enrichment (BIG-TREE), enables the rapid generation of clonal isogenic hPSC lines. It produces single-nucleotide editing efficiencies greater than 80% across multiple hPSC lines and allows for the efficient generation of loss-of-function hPSC lines and efficient multiplex editing of hPSCs at multiple independent loci.
BIG-TREE advances the implementation of base-editing technologies in hPSCs for use in developmental biology, disease modeling, drug screening, cell-based therapies and more.
• Isogenic cell line engineering
• Generating knockout cell lines
• Developmental biology
• Disease modeling
• Drug screening
• Regenerative medicine
Benefits and Advantages
• Greater than 80% efficiencies across multiple hPSC lines
• Allows for the precise and efficient base editing of hPSCs
• Efficient generation of loss-of-function hPSC lines
• Efficient editing of multiple loci and across several independent hPSC lines
• Bulk enrichment of base-edited cell populations including hPSCs
• Fast and efficient generation of clonal isogenic hPSC lines with homozygous and heterozygous single base pair edits
• Rapid engineering of isogenic hPSC lines
o Establishes in vitro models to assess pathogenic risk/disease mechanisms
• Allows for biallelic/multiplexed targeting without the need for sequential retargeting
• Compatible with off the shelf chemical transfection reagents
o Doesn’t require cloning of complex viral constructs/use of specialized cell transfection systems
• Can be used in conjunction with other base editing variants that have altered PAM specificities and editing windows
For more information about this opportunity, please see
For more information about the inventor(s) and their research, please see