Assembly of CRISPR ribonucleoproteins with biotinylated oligonucleotides via an RNA aptamer for precise gene editing

Jared Carlson-Stevermer, Amr A. Abdeen, Lucille Kohlenberg, Madelyn Goedland, Kaivalya Molugu, Meng Lou and Krishanu Saha ()
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Jared Carlson-Stevermer: University of Wisconsin-Madison
Amr A. Abdeen: University of Wisconsin-Madison
Lucille Kohlenberg: University of Wisconsin-Madison
Madelyn Goedland: University of Wisconsin-Madison
Kaivalya Molugu: University of Wisconsin-Madison
Meng Lou: University of Wisconsin-Madison
Krishanu Saha: University of Wisconsin-Madison

Nature Communications, 2017, vol. 8, issue 1, 1-13

Abstract: Abstract Writing specific DNA sequences into the human genome is challenging with non-viral gene-editing reagents, since most of the edited sequences contain various imprecise insertions or deletions. We developed a modular RNA aptamer-streptavidin strategy, termed S1mplex, to complex CRISPR-Cas9 ribonucleoproteins with a nucleic acid donor template, as well as other biotinylated molecules such as quantum dots. In human cells, tailored S1mplexes increase the ratio of precisely edited to imprecisely edited alleles up to 18-fold higher than standard gene-editing methods, and enrich cell populations containing multiplexed precise edits up to 42-fold. These advances with versatile, preassembled reagents could greatly reduce the time and cost of in vitro or ex vivo gene-editing applications in precision medicine and drug discovery and aid in the development of increased and serial dosing regimens for somatic gene editing in vivo.

Date: 2017
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DOI: 10.1038/s41467-017-01875-9

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