Testing multiplexed anti-ASFV CRISPR-Cas9 in reducing African swine fever virus.
Level 5 - mechanism / opinion, no new human data
Preclinical in vitro and animal challenge study (non-human).
PubMed 38563791 · doi:10.1128/spectrum.02164-23
What was done
Researchers developed a multiplexed CRISPR-Cas9 system targeting nine distinct loci in the African swine fever virus (ASFV) genome using Type II and Type III RNA polymerase-guided gRNA expression. After evaluating viral suppression in vitro, they generated a transgenic pig line constitutively expressing the CRISPR-Cas-gRNA system via germline genome editing and assessed antiviral efficacy and survival following in vivo ASFV challenge.
What was found
In vitro assays showed reduction in ASFV viral replication. Germline-edited transgenic pigs developed normally with continuous CRISPR-Cas-gRNA expression. Upon ASFV exposure, a subset of transgenic pigs exhibited latent viral replication and delayed infection; however, the CRISPR-Cas9-engineered pigs did not show any survival advantage. The abstract reports no numerical values, viral titers, or exact animal numbers.
Why it matters
This represents the first in vivo test of germline genome-edited animals engineered with an anti-ASFV CRISPR system. It demonstrates that in vitro suppression and delayed viral kinetics in vivo do not necessarily translate into clinical survival benefits against high-consequence viral pathogens.
Limits
The abstract provides no sample sizes (n) for in vitro experiments or animal challenge groups, and reports no quantitative metrics or statistical confidence intervals. The primary protective outcome (survival) was not achieved, and delayed infection was observed only in a subset of pigs.
Cited by
- supports George Church and Luhan Yang's group published research demonstrating the use of CRISPR to target and eliminate African swine fever virus DNA.