Lamb C, Pitre E, Ajufo S, Rigby C, Bisht K, Oade M. Quantification of influenza virus mini viral RNAs using Cas13. RNA. 2024 Oct 17:rna.080174.124
Influenza A virus (IAV) RNA synthesis produces full-length and deletion-containing RNA molecules, which include defective viral genomes (DVG) and mini viral RNAs (mvRNA). Sequencing approaches have shown that DVG and mvRNA species may be present during infection, and that they can vary in size, segment origin, and sequence. Moreover, a subset of aberrant RNA molecules can bind and activate host pathogen receptor retinoic acid-inducible gene I (RIG-I), leading to innate immune signaling and the expression of type I and III interferons. Measuring the kinetics and distribution of these immunostimulatory aberrant RNA sequences is important for understanding their function in IAV infection. Here, we explored if IAV mvRNA molecules can be detected and quantified using amplification-free, CRISPR-LbuCas13a-based detection. We show that CRISPR-LbuCas13a can be used to measure the copy numbers of specific mvRNAs in samples from infected tissue culture cells. However, to efficiently detect mvRNAs in other samples, promiscuous CRISPR guide RNAs are required that activate LbuCas13a in the presence of multiple mvRNA sequences. One crRNA was able to detect full-length IAV segment 5 without amplification, allowing it to be used for general IAV infection detection nasopharyngeal swabs. Using CRISPR-LbuCas13a, we confirm that mvRNAs are present in ferret upper and lower respiratory tract tissue, as well as clinical nasopharyngeal swab extracts of hospitalized patients. Overall, CRISPR-LbuCas13a-based RNA detection is a useful tool for studying deletion-containing viral RNAs and it complements existing amplification-based approaches.
See Also:
Latest articles in those days:
- Characterisation of a human monoclonal antibody targeting a conserved epitope at the base of the HA head of influenza A(H3N2) virus 5 hours ago
- An outbreak report of Influenza A(H3) among residents of a large nursing home in northern Italy: attack rates, clinical features and outcomes 18 hours ago
- Susceptibility of calves fed unpasteurized milk from cows infected with highly pathogenic avian influenza H5N1 18 hours ago
- Functional mapping of IFITM3 resolves its antiviral core and deciphers human influenza susceptibility 18 hours ago
- Near real-time data on the human neutralizing antibody landscape to influenza virus as of early 2026 to inform vaccine-strain selection 18 hours ago
[Go Top] [Close Window]


