Details

Autor(en) / Beteiligte

• Ashraf, Muhammad Usman
• Salman, Hafiz Muhammad
• Khalid, Muhammad Farhan
• Khan, Muhammad Haider Farooq
• Anwar, Saima
• Afzal, Samia
• Idrees, Muhammad
• Chaudhary, Safee Ullah

Titel

CRISPR-Cas13a mediated targeting of hepatitis C virus internal-ribosomal entry site (IRES) as an effective antiviral strategy

Ist Teil von

• Biomedicine & pharmacotherapy, 2021-04, Vol.136, p.111239-111239, Article 111239

Ort / Verlag

France: Elsevier Masson SAS

Erscheinungsjahr

2021

Quelle

Elsevier ScienceDirect Journals Complete

Beschreibungen/Notizen

• [Display omitted] •CRISPR effector Cas13a can be reprogrammed to target ssRNA virus in a eukaryotic system.•Cas13a is effective at targeting hepatitis C virus in cell culture.•Several of potential Cas13a target sites are present in HCV-IRES.•Optimal Cas13a target sites are identified via computational analysis.•Cas13a-based targeting of HCV-IRES can be used to develop broad genotypic therapeutics. Hepatitis C is an inflammatory liver disease caused by the single-stranded RNA (ssRNA) hepatitis C virus (HCV). The genetic diversity of the virus and quasispecies produced during replication have resulted in viral resistance to direct-acting antivirals (DAAs) as well as impediments in vaccine development. The recent adaptation of CRISPR-Cas as an alternative antiviral approach has demonstrated degradation of viral nucleic acids in eukaryotes. In particular, the CRISPR-effector Cas13 enzyme has been shown to target ssRNA viruses effectively. In this work, we have employed Cas13a to knockdown HCV in mammalian cells. Using a computational screen, we identified several potential Cas13a target sites within highly conserved regions of the HCV internal ribosomal entry site (IRES). Our results demonstrate significant inhibition of HCV replication as well as translation in huh-7.5 cells with minimal effects on cell viability. These findings were validated using a multi-modality approach involving qRT-PCR, luciferase assay, and MTT cell viability assay. In conclusion, the CRISPR-Cas13a system efficiently targets HCV in vitro, suggesting its potential as a programmable therapeutic antiviral strategy.