Data from PNA-Assisted DNAzymes to Cleave Double-Stranded DNA for Genetic Engineering with High Sequence Fidelity

Dataset · Tue Jun 22 00:00:00 EDT 2021

DOI:https://doi.org/10.13012/B2IDB-1858504_V1· OSTI ID:3014136

Lyu, Mingkuan ^[1]; Kong, Linggen ^[2]; Yang, Zhenglin ^[3]; Wu, Yuting ^[4]; McGhee, Claire E. ^[4]; Lu, Yi ^[5]

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1. Department of Chemistry, University of Illinois at Urbana−Champaign, Urbana, Illinois 61801, United States; Center for Advanced Bioenergy and Bioproducts Innovation (CABBI), Urbana, IL (United States)
2. Center for Biophysics and Quantitative Biology, University of Illinois at Urbana−Champaign, Urbana, Illinois 61801, United States
3. Department of Biochemistry, University of Illinois at Urbana−Champaign, Urbana, Illinois 61801, United States
4. Department of Chemistry, University of Illinois at Urbana−Champaign, Urbana, Illinois 61801, United States
5. Department of Chemistry, University of Illinois at Urbana−Champaign, Urbana, Illinois 61801, United States; Center for Biophysics and Quantitative Biology, University of Illinois at Urbana−Champaign, Urbana, Illinois 61801, United States; Department of Biochemistry, University of Illinois at Urbana−Champaign, Urbana, Illinois 61801, United States; Carl R. Woese Institute for Genomic Biology, University of Illinois at Urbana−Champaign, Urbana, Illinois 61801, United States; Center for Advanced Bioenergy and Bioproducts Innovation (CABBI), Urbana, IL (United States)

DNAzymes have been widely used in many sensing and imaging applications but have rarely been used for genetic engineering since their discovery in 1994, because their substrate scope is mostly limited to single-stranded DNA or RNA, whereas genetic information is stored mostly in double-stranded DNA (dsDNA). To overcome this major limitation, we herein report peptide nucleic acid (PNA)-assisted double-stranded DNA nicking by DNAzymes (PANDA) as the first example to expand DNAzyme activity toward dsDNA. We show that PANDA is programmable in efficiently nicking or causing double strand breaks on target dsDNA, which mimics protein nucleases and can act as restriction enzymes in molecular cloning. In addition to being much smaller than protein enzymes, PANDA has a higher sequence fidelity compared with CRISPR/Cas under the condition we tested, demonstrating its potential as a novel alternative tool for genetic engineering and other biochemical applications.

• Research Organization: Center for Advanced Bioenergy and Bioproducts Innovation (CABBI), Urbana, IL (United States); University of Illinois Urbana-Champaign
• Sponsoring Organization: U.S. Department of Energy (DOE)
• DOE Contract Number: SC0018420
• OSTI ID: 3014136
• Country of Publication: United States
• Language: English

Related Subjects

Conversion
Genome Engineering
Genomics