2015 Synthetic Biology: Engineering, Evolution & Design (SEED)
Synthetic nucleic acid nanostructures regulate native triplex and CRISPR/Cas genomic recombination, restriction, and repair potentials
Author
CONFERENCE:
AIChE Society for Biological Engineering,
Synthetic Biology: Engineering, Evolution, and Design (SEED) 2015 Conference,
Boston, MA, USA June 10-13, 2015
SUBMISSION HEREIN:
Conference Abstract for Speaker Slot/Platform Talk
SUITABLE CONFERENCE TRACK:
DNA and RNA Based Synthetic Biology
AUTHOR:
Faisal Reza, Ph.D.
CORRESPONDING AUTHOR:
Email: faisal.reza@yale.edu
Synthetic nucleic acid nanostructures regulate native triplex and CRISPR/Cas genomic recombination, restriction, and repair potentials
Faisal Reza, Ph.D.â?
â? Yale University, School of Medicine, Department of Therapeutic Radiology, New Haven, CT, USA
*Address all correspondence to:
Faisal Reza; Yale University, School of Medicine, Department of Therapeutic Radiology, 333 Cedar Street, HRT 316, #208040, New
Haven, CT, 06520-8040, USA; Email: faisal.reza@yale.edu.
Synthetic oligo- and peptide- nucleic acid nanostructures have been more predictably designed and externally delivered into the intracellular milieu. These deployed technologies interact with, and influence, the cytoplasmic and nuclear molecular machinery in order to regulate potentials involved in genomic targeting and editing. It is demonstrated that synthetic nucleic acid nanostructures composed of various nucleobase and backbone modifications can regulate the genomic recombination rate, the sequence-specific restriction of a locus, and the endogenous repair pathways. The formation of a triplex nanostructure, by exogenously introduced PNA molecules with the duplex chromosomal and episomal DNA, is shown to elevate the cellâ??s targeted recombination potential. Alternatively, the formation of a
RNA-guided CRISPR/Cas nanostructure, again by exogenously introduced RNA molecules with Cas nucleases, is shown to initiate the cellâ??s targeted double-strand restriction potential. Recombinagenic donor DNA molecules co-opt these elevated recombination or initiated restriction potentials to form competing nanostructures that act as homology-dependent templates, sans edits to be introduced, thus potentiating repair. Safety and efficacy of these nanostructures is achieved by leveraging the performance profile of the cell's own endogenous recombination, restriction, and repair machineries in concert with these sequence-specific and localizing-in-tandem molecules. Progenitor cells drugged with designed molecules, and primed with chemical cell modulators, safely and effectively redesign the genome, which are then propagated to cellular progeny. These molecular technologies are developed to remediate the underlying genomic causes of monogenic human diseases, engineer living genetic codes, improve crop characteristics, and defend against outbreaks, and thus has well-positioned technology profiles in the synthetic biology of healthcare, biotechnology, agrotechnology, and national security.