Page 1 - J. Chromatin remodeling and epigenetics

Development Of aMTD-Cas9/sgRNA To Overcome The Current
Limitations Of The CRISPR-Cas9 System Using TSDT

Yongchan Choi, Heeseop Yoo, Jinwook Yang, Youngsil Choi and Daewoong Jo
Cas9 Team, Cellivery R&D Institute, Cellivery Therapeutics, Inc., Seoul 03929, Korea.
BACKGROUND AIM
CRISPR-Cas9 system is a highly specific genome modification tool, Providing a novel cell-permeable Cas9 system by making aMTD-
which is composed of Cas9 nuclease and sgRNA. In spite of their
therapeutic potential in treating genetic diseases, however, the Cas9/sgRNA ribonucleoprotein (RNP) capable of direct cell delivery,
as a therapeutic gene-editing module that is practical to target
clinical applicability of CRISPR/Cas9 system is limited by low
cell/tissue delivery efficiency. human genetic diseases via genome modification.
METHODS
Cell-permeable (CP) Cas9 recombinant protein was developed by fusing sequence-optimized hydrophobic cell-penetrating peptide (CPP),
namely advanced macromolecule transduction domain (aMTD), to deliver Cas9 directly into cells and tissues. GM06214 cells were treated with
FITC-labeled His-NLS-aMTD323-Cas9 for 3 hrs. After incubation, cells were harvested and analyzed by flow cytometry.

RESULTS

CONCLUSION REFERENCES Contact information
Chung et al. (2020) Science Advances, 6: eaba 1193 Minyong Jung
By using additional carrier such as Lim et al. (2013) Clinical Cancer Research, 19: 680-690
Lipofectamine, sgRNA showed gene-editing New Drug & Business Development
activity with aMTD-Cas9. In cell experiments, Lim et al. (2013) Biomaterials, 34: 6261-6271
aMTD-Cas9 / sgRNA ribo nucleoprotein Lim et al. (2012) Molecular Therapy, 20: 1540-1549 Cellivery Therapeutics, Inc.
(RNP) alone showed activity without jungmy@cellivery.com
additional treatment with sgRNA. Jo et al. (2005) Nature Medicine, 11: 892-898
Jo et al. (2001) Nature Biotechnology, 19: 929-933 +82-2-3151-8900