Cholesterol-functionalized DNA/RNA heteroduplexes cross the blood–brain barrier and knock down genes in the rodent CNS

Tetsuya Nagata, Chrissa A. Dwyer, Kie Yoshida-Tanaka, Kensuke Ihara, Masaki Ohyagi, Hidetoshi Kaburagi, Haruka Miyata, Satoe Ebihara, Kotaro Yoshioka, Takashi Ishii, Kanjiro Miyata, Kenichi Miyata, Berit Powers, Tomoko Igari, Syunsuke Yamamoto, Naoto Arimura, Hideki Hirabayashi, Toshiki Uchihara, Rintaro Iwata Hara, Takeshi WadaC. Frank Bennett, Punit P. Seth, Frank Rigo, Takanori Yokota

Research output: Contribution to journalArticlepeer-review

52 Citations (Scopus)


Achieving regulation of endogenous gene expression in the central nervous system (CNS) with antisense oligonucleotides (ASOs) administered systemically would facilitate the development of ASO-based therapies for neurological diseases. We demonstrate that DNA/RNA heteroduplex oligonucleotides (HDOs) conjugated to cholesterol or α-tocopherol at the 5′ end of the RNA strand reach the CNS after subcutaneous or intravenous administration in mice and rats. The HDOs distribute throughout the brain, spinal cord and peripheral tissues and suppress the expression of four target genes by up to 90% in the CNS, whereas single-stranded ASOs conjugated to cholesterol have limited activity. Gene knockdown was observed in major CNS cell types and was greatest in neurons and microglial cells. Side effects, such as thrombocytopenia and focal brain necrosis, were limited by using subcutaneous delivery or by dividing intravenous injections. By crossing the blood–brain barrier more effectively, cholesterol-conjugated HDOs may overcome the limited efficacy of ASOs targeting the CNS without requiring intrathecal administration.

Original languageEnglish
Pages (from-to)1529-1536
Number of pages8
JournalNature Biotechnology
Issue number12
Publication statusPublished - 2021 Dec
Externally publishedYes

ASJC Scopus subject areas

  • Biotechnology
  • Bioengineering
  • Applied Microbiology and Biotechnology
  • Molecular Medicine
  • Biomedical Engineering


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