Case ID: M20-204L^

Published: 2021-03-23 15:18:12

Last Updated: 1670405556


Inventor(s)

David Brafman
Xiao Wang
Nicholas Brookhouser
Stefan Tekel
Kylie Standage-Beier

Technology categories

Genomic Assays/Reagents/ToolsLife Science (All LS Techs)

Licensing Contacts

Jovan Heusser
Director of BD LS
[email protected]

Base-Edited Isogenic hPSC Line Generation (BIG-TREE)

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.

 

Potential Applications

•       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

Brookhouseer et al – Stem Cell Reports – 2020

For more information about the inventor(s) and their research, please see

Dr. Brafman’s departmental webpage

Dr. Wang’s laboratory webpage