Repurposing CRISPR/Cas to Discover SARS-CoV-2 Detecting and Neutralizing Aptamers.

RNA aptamers provide useful biological probes and therapeutic agents. New methodologies to screen RNA aptamers will be valuable by complementing the traditional Systematic Evolution of Ligands by Exponential Enrichment (SELEX). Meanwhile, repurposing clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR associated systems (Cas) has expanded their utility far beyond their native nuclease function. Here, CRISmers, a CRISPR/Cas-based novel screening system for RNA aptamers based on binding to a chosen protein of interest in a cellular context, is presented. Using CRISmers, aptamers are identified specifically targeting the receptor binding domain (RBD) of the spike glycoprotein of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). Two aptamer leads enable sensitive detection and potent neutralization of SARS-CoV-2 Delta and Omicron variants in vitro. Intranasal administration of one aptamer, further modified with 2'-fluoro pyrimidines (2'-F), 2'-O-methyl purines (2'-O), and conjugation with both cholesterol and polyethylene glycol of 40 kDa (PEG40K), achieves effective prophylactic and therapeutic antiviral activity against live Omicron BA.2 variants in vivo. The study concludes by demonstrating the robustness, consistency, and potential broad utility of CRISmers using two newly identified aptamers but switching CRISPR, selection marker, and host species.

Three new species of Macrostomum (Platyhelminthes, Macrostomorpha) from China and Australia, with notes on taxonomy and phylogenetics.

In this paper, three species of the macrostomid free-living flatworm genus Macrostomum are described. Two species, Macrostomumlittorale Wang & Shi, sp. nov. and M.shekouense Wang & Shi, sp. nov., were collected from coastal water at Shenzhen, Guangdong Province, China. One species, M.brandi Wang & Shi, sp. nov., was collected from Perth, Western Australia and Queenscliff, Victoria, Australia. Macrostomumlittorale sp. nov. differs from congeneric species within the genus in the length of the stylet, diameter of stylet opening, and the bend of the stylet. Macrostomumshekouense sp. nov. and M.brandi sp. nov. differ from similar species within the genus in the stylet morphology, position of the female antrum and developing eggs, or presence or absence of the false seminal vesicle. Phylogenetic analysis based on cytochrome c oxidase subunit I (COI) gene shows that M.littorale sp. nov. and M.hystrix are sister clades on two well-separated branch, M.shekouense sp. nov. and M.brandi sp. nov. are sister clades on two well-separated branches. Accordingly, both morphological and molecular evidence support M.littorale sp. nov., M.shekouense sp. nov., and M.brandi sp. nov. as three new species.

Two New Brackish-Water Species of Macrostomum (Platyhelminthes: Macrostomorpha) from China and Their Phylogenetic Positions.

In this paper, two new brackish-water species of the macrostomid turbellarian genus Macrostomum, Macrostomum pseudosinense sp. nov. and Macrostomum taurinum sp. nov., collected from coastal water at Shenzhen, Guangdong Province, China, are described based on morphological, histological, and molecular phylogenetic analyses. Macrostomum pseudosinense sp. nov. differs from similar species within the genus in the length of the stylet (152 ± 15.0 µm), diameter of stylet opening (20 ± 4.0 µm proximally; 7 ± 0.5 µm distally), two bends of the stylet, and the non-spiral end of the stylet. Macrostomum taurinum sp. nov. differs from its congeners in the length of the stylet (81 ± 7.4 µm), the stylet bending position and angle (50% and 60°), diameter of stylet proximal opening (15 ± 3.0 µm), sperm with bristles and brush, and the smooth-walled ovaries. Phylogenetic analyses inferred from nuclear 18S and 28S rRNA genes support the establishments of these two new species. In addition, reciprocal mating behavior of M. pseudosinense sp. nov. was observed and documented.

Macrostomum shenda and M. spiriger, two new brackish-water species of Macrostomum (Platyhelminthes: Macrostomorpha) from China.

In this study, two new brackish-water species of Macrostomum (M. shenda n. sp. and M. spiriger n. sp.) collected from Shenzhen, China, were described based on morphological, histological, and molecular phylogenetic analyses. Macrostomum shenda n. sp. differs from its congeners in the oblique and non-thickening distal opening of its narrow funnel-shaped stylet. In addition, its sperm have neither bristles nor brush, and are surrounded by an undulating membrane in the mid-body region. In M. spiriger n. sp., the stylet is spirally twisted. Its reproductive apparatus has a seminal bursal pore opening exteriorly. Results of the 18S and 28S rDNA phylogenetic analyses also support the establishments of these two new species. Moreover, the 18S and 28S rDNA sequences of some species within Macrostomum in previous studies have been revised to avoid ambiguity, while Macrostomum dongyuanensis Wang Sun, 2015 was re-identified as a new record of M. quiritium Kolasa, 1973 from China.