NP-RNA interaction screen

interaction in bacteria

(Woo Cho 2019 pmid: 31594639) screen in bacteria - cell-based LacZ translation inhibition assay that probes for specific interactions between RNA-binding proteins and their target sequences [23,24]. In this assay, putative target sequences are placed upstream of a LacZ reporter gene; if an RNA binding protein expressed in the cell recognizes and binds to a target sequence, LacZ translation is inhibited. Moreover, the strength of the interaction between the RNA binding protein and its target sequence is directly proportional to the degree of inhibition of LacZ translation.

In mammalian cell culture we would screen for general Cov2 inhibitors.

(Xie Zhang 2020 pmid: 32289263) reporter virus - everse genetic system for SARS-CoV-2. Seven complimentary DNA (cDNA) fragments spanning the SARSCoV-2 genome were assembled into a full-genome cDNA. Compared with a clinical isolate, the infectiousclone-derived SARS-CoV-2 (icSARS-CoV-2) exhibited similar plaque morphology, viral RNA profile, and replication kinetics.

We generated a stable mNeonGreen SARS-CoV-2 (icSARS-CoV-2-mNG) by introducing this reporter gene into ORF7 of the viral genome.

the mNG-virus-based assay could be automated in a high-throughput format to screen compounds against viral replication.

transfection, mutated s-protein - 2 color system n- to go to the virus and reverse transcriptase for background