TY  - JOUR
AB  - High-capacity adenoviral vectors (HCAdVs) devoid of all coding genes are powerful tools to deliver large DNA cargos into cells. Here HCAdVs were designed to deliver a multiplexed complete CRISPR/Cas9 nuclease system or a complete pair of transcription activator-like effector nucleases (TALENs) directed against the hepatitis B virus (HBV) genome. HBV, which remains a serious global health burden, forms covalently closed circular DNA (cccDNA) as a persistent DNA species in infected cells. This cccDNA promotes the chronic carrier status, and it represents a major hurdle in the treatment of chronic HBV infection. To date, only one study demonstrated viral delivery of a CRISPR/Cas9 system and a single guide RNA (gRNA) directed against HBV by adeno-associated viral (AAV) vectors. The advancement of this study is the co-delivery of multiple gRNA expression cassettes along with the Cas9 expression cassette in one HCAdV. Treatment of HBV infection models resulted in a significant reduction of HBV antigen production and the introduction of mutations into the HBV genome. In the transduction experiments, the HBV genome, including the HBV cccDNA, was degraded by the CRISPR/Cas9 system. In contrast, the combination of two parts of a TALEN pair in one vector could not be proven to yield an active system. In conclusion, we successfully delivered the CRISPR/Cas9 system containing three gRNAs using HCAdV, and we demonstrated its antiviral effect.
AU  - Schiwon, M.*
AU  - Ehrke-Schulz, E.*
AU  - Oswald, A.
AU  - Bergmann, T.*
AU  - Michler, T.
AU  - Protzer, U.
AU  - Ehrhardt, A.*
C1  - 53832
C2  - 45069
CY  - 50 Hampshire St, Floor 5, Cambridge, Ma 02139 Usa
SP  - 242-253
TI  - One-vector system for multiplexed CRISPR/Cas9 against hepatitis B virus cccDNA utilizing high-capacity adenoviral vectors.
JO  - Mol. Ther. Nucleic Acids
VL  - 12
PB  - Cell Press
PY  - 2018
SN  - 2162-2531
ER  - 

TY  - JOUR
AB  - Changes in lifestyle and environmental conditions give rise to an increasing prevalence of liver and lung fibrosis, and both have a poor prognosis. Promising results have been reported for recombinant angiotensin-converting enzyme 2 ( ACE2) protein administration in experimental liver and lung fibrosis. However, the full potential of ACE2 may be achieved by localized translation of a membrane-anchored form. For this purpose, we advanced the latest RNA technology for liver-and lung-targeted ACE2 translation. We demonstrated in vitro that transfection with ACE2 chemically modified messenger RNA (cmRNA) leads to robust translation of fully matured, membrane-anchored ACE2 protein. In a second step, we designed eight modified ACE2 cmRNA sequences and identified a lead sequence for in vivo application. Finally, formulation of this ACE2 cmRNA in tailor-made lipidoid nanoparticles and in lipid nanoparticles led to liver-and lung-targeted translation of significant amounts of ACE2 protein, respectively. In summary, we provide evidence that RNA transcript therapy (RTT) is a promising approach for ACE2-based treatment of liver and lung fibrosis to be tested in fibrotic disease models.
AU  - Schrom, E.*
AU  - Huber, M.*
AU  - Aneja, M.K.*
AU  - Dohmen, C.*
AU  - Emrich, D.*
AU  - Geiger, J.*
AU  - Hasenpusch, G.*
AU  - Herrmann-Janson, A.*
AU  - Kretzschmann, V.*
AU  - Mykhailyk, O.*
AU  - Pasewald, T.*
AU  - Oak, P.
AU  - Hilgendorff, A.
AU  - Wohlleber, D.*
AU  - Hoymann, H.*
AU  - Schaudien, D.*
AU  - Plank, C.*
AU  - Rudolph, C.*
AU  - Kubisch-Dohmen, R.*
C1  - 51213
C2  - 42922
CY  - Cambridge
SP  - 350-365
TI  - Translation of angiotensin-converting enzyme 2 upon liver- and lung-targeted delivery of optimized chemically modified mRNA.
JO  - Mol. Ther. Nucleic Acids
VL  - 7
PB  - Cell Press
PY  - 2017
SN  - 2162-2531
ER  -