In vivo delivery of transcription factors with multifunctional oligonucleotides.
Therapeutics based on transcription factors have the potential to revolutionize medicine but have had limited clinical success as a consequence of delivery problems. The delivery of transcription factors is challenging because it requires the development of a delivery vehicle that can complex transcription factors, target cells and stimulate endosomal disruption, with minimal toxicity. Here, we present a multifunctional oligonucleotide, termed DARTs (DNA assembled recombinant transcription factors), which can deliver transcription factors with high efficiency in vivo. DARTs are composed of an oligonucleotide that contains a transcription-factor-binding sequence and hydrophobic membrane-disruptive chains that are masked by acid-cleavable galactose residues. DARTs have a unique molecular architecture, which allows them to bind transcription factors, trigger endocytosis in hepatocytes, and stimulate endosomal disruption. The DARTs have enhanced uptake in hepatocytes as a result of their galactose residues and can disrupt endosomes efficiently with minimal toxicity, because unmasking of their hydrophobic domains selectively occurs in the acidic environment of the endosome. We show that DARTs can deliver the transcription factor nuclear erythroid 2-related factor 2 (Nrf2) to the liver, catalyse the transcription of Nrf2 downstream genes, and rescue mice from acetaminophen-induced liver injury.
1] Department of Bioengineering, University of California, Hearst Memorial Mining Building, Room 284 Berkeley, California 94720, USA [2] UC Berkeley and UCSF Joint Graduate Program in Bioengineering, Berkeley/San Francisco, USA.
Kunwoo
K
Lee
Department of Bioengineering, University of California, Hearst Memorial Mining Building, Room 284 Berkeley, California 94720, USA.
Mohammad
M
Rafi
Department of Bioengineering, University of California, Hearst Memorial Mining Building, Room 284 Berkeley, California 94720, USA.
Xiaojian
X
Wang
Department of Bioengineering, University of California, Hearst Memorial Mining Building, Room 284 Berkeley, California 94720, USA.
Kiana
K
Aran
Department of Bioengineering, University of California, Hearst Memorial Mining Building, Room 284 Berkeley, California 94720, USA.
Xuli
X
Feng
Department of Bioengineering, University of California, Hearst Memorial Mining Building, Room 284 Berkeley, California 94720, USA.
Carlo
C
Lo Sterzo
Department of Bioengineering, University of California, Hearst Memorial Mining Building, Room 284 Berkeley, California 94720, USA.
Richard
R
Tang
Department of Bioengineering, University of California, Hearst Memorial Mining Building, Room 284 Berkeley, California 94720, USA.
Nithya
N
Lingampalli
Department of Bioengineering, University of California, Hearst Memorial Mining Building, Room 284 Berkeley, California 94720, USA.
Hyun Jin
HJ
Kim
1] Department of Bioengineering, University of California, Hearst Memorial Mining Building, Room 284 Berkeley, California 94720, USA [2] UC Berkeley and UCSF Joint Graduate Program in Bioengineering, Berkeley/San Francisco, USA.
Niren
N
Murthy
D000410:Alanine Transaminase / Q000378:metabolism; D000818:Animals; D004247:DNA / Q000737:chemistry*; D016503:Drug Delivery Systems; D011992:Endosomes / Q000378:metabolism; D056945:Hep G2 Cells; D022781:Hepatocytes / Q000166:cytology; D006801:Humans; D006863:Hydrogen-Ion Concentration; D006868:Hydrolysis; D008099:Liver / Q000378:metabolism; D051379:Mice; D051267:NF-E2-Related Factor 2 / Q000378:metabolism; D009841:Oligonucleotides / Q000737:chemistry*; D018384:Oxidative Stress; D013050:Spectrometry, Fluorescence; D014018:Tissue Distribution; D014157:Transcription Factors / Q000378:metabolism*
D051267:NF-E2-Related Factor 2; C495636:Nfe2l2 protein, mouse; D009841:Oligonucleotides; D014157:Transcription Factors; D004247:DNA; D000410:Alanine Transaminase