Establishment of a pig CRISPR/Cas9 knockout library for functional gene screening in pig cells

Abstract

Supporting information updated on 18 November 2021, after first online publication.

A Nectin1 Mutant Mouse Model Is Resistant to Pseudorabies Virus Infection

“…Second, other reported gene targets, such as PVR, which may play a key role in PRV cell entry, can be investigated in animals with respect their antiviral ability in vivo [ 16 , 36 ]. Third, the recently developed CRISPR library can effectively screen host genes essential for viral infection [ 37 , 38 ]. A genome-wide screen has been performed for some alphaherpesviruses including PRV and Bovine Herpes Virus Type 1 (BHV-1) and revealed more potential gene targets, which are promising resources facilitating antiviral study [ 39 , 40 ].…”

Advances in CRISPR-Based Functional Genomics and Nucleic Acid Detection in Pigs

“…related to Japanese encephalitis virus (JEV) infection ( Zhao et al, 2020 ). Several reports have identified multiple host factors required for the entry of other viruses and toxins in pigs and humans by using the CRISPR screening strategy ( Hölper et al, 2021 ; Luo et al, 2021 ; Sun et al, 2021 ; Yu et al, 2021 ; Zhou et al, 2021 ).…”

“…Host sphingomyelin synthase 1 (SMS1) was also found to be involved in pseudorabies virus (PRV) infection when the gD-mediated entry pathway was blocked ( Hölper et al, 2021 ). In addition, HBEGF (heparin-binding EGF-like growth factor), DPH1-5 (diphthamide biosynthesis 1–5), DNAJC24 (Hsp40 member C24), and ZBTB17 were determined as diphtheria toxin (DT) receptors ( Yu et al, 2021 ). These are the key factors involved in the biosynthesis of diphthamide, which serves as the molecular target for DT ( Yu et al, 2021 ).…”

“…In addition, HBEGF (heparin-binding EGF-like growth factor), DPH1-5 (diphthamide biosynthesis 1–5), DNAJC24 (Hsp40 member C24), and ZBTB17 were determined as diphtheria toxin (DT) receptors ( Yu et al, 2021 ). These are the key factors involved in the biosynthesis of diphthamide, which serves as the molecular target for DT ( Yu et al, 2021 ). These data demonstrate that CRISPR screening strategy is a powerful tool for functional genome in livestock.…”

Editorial: CRISPR and alternative approaches

“…shared an exciting development of a CRISPR knockout library to perform functional genomics screenings in pig cells, and the generation of transgenic pigs through a CRISPR‐based approach, which brings transformative gene‐editing technology into this important animal. [ 4 ,5] Habu et al. revealed a new safe harbor locus for mammalian cell gene‐editing, leading to new possibilities for both basic and translational research.…”