A Crucial Role of ACBD3 Required for Coxsackievirus Infection in Animal Model Developed by AAV-Mediated CRISPR Genome Editing Technique

Abstract

Genetic screens using CRISPR/Cas9 have been exploited to discover host–virus interactions. These screens have identified viral dependencies on host proteins during their life cycle and potential antiviral strategies. The acyl-CoA binding domain containing 3 (ACBD3) was identified as an essential host factor for the Coxsackievirus B3 (CVB3) infection. Other groups have also investigated the role of ACBD3 as a host factor for diverse enteroviruses in cultured cells. However, it has not been tested if ACBD3 is required in the animal model of CVB3 infection. Owing to embryonic lethality, conventional knockout mice were not available for in vivo study. As an alternative approach, we used adeno-associated virus (AAV)-mediated CRISPR genome editing to generate mice that lacked ACBD3 within the pancreas, the major target organ for CVB3. Delivery of sgRNAs using self-complementary (sc) AAV8 efficiently induced a loss-of-function mutation in the pancreas of the Cas9 knock-in mice. Loss of ACBD3 in the pancreas resulted in a 100-fold reduction in the CVB3 titer within the pancreas and a noticeable reduction in viral protein expression. These results indicate a crucial function of ACBD3 in CVB3 infection in vivo. AAV-mediated CRISPR genome editing may be applicable to many in vivo studies on the virus–host interaction and identify a novel target for antiviral therapeutics.

Post-genomic era in agriculture and veterinary science: successful and proposed application of genetic targeting technologies

“…Recently, a genome-wide screen using CRISPR/Cas9 was conducted to identify host factors that may potentially participate in the replication of certain viruses, such as SARS-CoV-2 ( 90 ), HIV ( 91 ), arthritic alphaviruses ( 92 ), Coxsackie virus ( 93 ), Venezuelan equine encephalitis virus (VEEV) ( 94 ) and influenza virus ( 95 ). For the influenza virus, it was demonstrated that inactivation of specific host genes, such as DOCK5, Annexin-A1, IFIT2, IRF7 and ZDHHC22, can induce protection against cell death during infection ( 96 – 100 ).…”

Latest Advances of Virology Research Using CRISPR/Cas9-Based Gene-Editing Technology and Its Application to Vaccine Development

“…Recently, genome-wide CRISPR/Cas9 screening has been used to identify host factors that are potentially involved in the replications of some viruses, such as the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) [ 96 ], HIV [ 97 ], arthritogenic alphaviruses [ 98 ], Coxsackievirus [ 99 ], Venezuelan equine encephalitis virus (VEEV) [ 100 ], and influenza virus [ 101 ]. For the influenza virus, it has been demonstrated that inactivation of the specific host genes, such as DOCK5, Annexin-A1, IFIT2, IRF7, and ZDHHC22, can induce protection against cell death by influenza virus infection [ 102 , 103 , 104 , 105 , 106 ].…”