An enhanced broad-spectrum peptide inhibits Omicron variants in vivo

. 2024 Feb 9;5(2):101418. doi: 10.1016/j.xcrm.2024.101418

REAGENT or RESOURCE	SOURCE	IDENTIFIER
Antibodies

Rabbit anti-SARS-CoV-2 N protein	In house	N/A
goat anti-rabbit Alexa Fluor 488-conjugated IgG (H + L)	Thermo Fisher	Cat#: A-11008; RRID: AB_143165

Bacterial and virus strains

BL21	In house	N/A
SARS-CoV-2 Alpha	Shuai et al. 2022³⁹	GenBank: OM212469
SARS-CoV-2 Omicron BA.2.12.1	Chon et al. 2023⁴⁰	EPI_ISL_13777659
SARS-CoV-2 Omicron BA.5	Chon et al. 2023⁴⁰	EPI_ISL_13777658
SARS-CoV-2 Omicron EG.5.1	Isolated from nasopharyngeal swab of a laboratory confirmed patient	EPL_ISL_18461518

Chemicals, peptides, and recombinant proteins

A1	This paper	N/A
HR2m	This paper	N/A
HR1	This paper	N/A
HR2m-Chol	This paper	N/A
A1-Chol	This paper	N/A
A1L35HR2m-Chol	This paper	N/A
A1L35HR2m	This paper	N/A
A1L35HR2mdCdN	This paper	N/A
A3L35HR2m	This paper	N/A
A7L35HR2m	This paper	N/A
isopropyl 1-thio-β-D-galactopyranoside	Sangon Biotech	Cat#: A600168-0005
Vigofect	Vigorous Biotech	Cat#: T001
EZ-Link NHS-PEG12-Biotin	Thermo Fisher Scientific	Cat#: 21313
Dulbecco’s Modified Eagle’s Medium	Gibco	Cat#: 11965-092
Fetal Bovine Serum	Gibco	Cat#: 16140071
G418	MCE	Cat#: HY-17561
Phosphate buffered saline	Gibco	Cat#: 10010-023
Penicillin/Streptomycin	Gibco	Cat#: 15140122
0.25% Trypsin-EDTA	Gibco	Cat#: 25200-072

Critical commercial assays

Luciferase assay system	Promega	Cat#: E1501
Cell Counting Kit-8	Dojindo	Cat#: CK04
Alanine aminotransferase Assay Kit	NJJCBIO	Cat#: C009-2-1
Aspartate aminotransferase Assay Kit	NJJCBIO	Cat#: C010-2-1
γ- Glutamyl transferase Assay Kit	NJJCBIO	Cat#: C017-2-1
creatinine (Cr) Assay kit	NJJCBIO	Cat#: C011-2-1
RNeasy mini kit	Qiagen	Cat#: 74106
One Step TB Green® PrimeScript™ RT-PCR Kit II (Perfect Real Time)	Takara	Cat#: RR086A

Experimental models: Cell lines

293T	ATCC	CRL-3216
Caco-2	Cai et al., 2020²⁹	N/A
Calu-3	Xia et al., 2020¹⁴	N/A
VeroE6-TMPRSS2	Japanese Collection of Research Bioresources (JCRB) Cell Bank	JCRB1819

Experimental models: Organisms/strains

Balb/c mice	Laboratory Animal Resources Center of Westlake University	N/A
K18-hACE2 C57BL/6J mice	The Jackson Laboratory	Strain#: 034860

Oligonucleotides

SARS-CoV-2 RdRp forward sequence: 5′-CGCATACAGTCTTRCAGGCT-3′	Integrated DNA Technologies	N/A
SARS-CoV-2 RdRp reverse sequence: 5′-GTGTGATGTTGAWATGACATGGTC-3′	Integrated DNA Technologies	N/A
Housekeeping gene mouse beta-actin sequence forward: 5′- ACGGCCAGGTCATCACTATTG-3′	Integrated DNA Technologies	N/A
Housekeeping gene mouse beta-actin reverse sequence: 5′- CAAGAAGGAAGGCTGGAAAAG-3′	Integrated DNA Technologies	N/A

Recombinant DNA

pcDNA3.1-SARS-CoV-S	Xia et al., 2019¹⁴	GenBank: ABD72979.1
pcDNA3.1-MERS-CoV-S	MiaoLingBio	N/A
pcDNA3.1-SARS-CoV-2-S	Xia et al., 2020²⁰	GenBank: QHD43416
pcDNA3.1-SARS-CoV-2 D614G S	This paper	N/A
pcDNA3.1-SARS-CoV-2 Beta S	This paper	N/A
pcDNA3.1-SARS-CoV-2 Delta S	This paper	N/A
pcDNA3.1-SARS-CoV-2 Omicron BA.1 S	Genewiz	N/A
pcDNA3.1-SARS-CoV-2 Omicron XBB S	This paper	N/A
pcDNA3.1-SARS-CoV-2 Omicron BQ.1.1 S	This paper	N/A
pcDNA3.1-EGFP	This paper	N/A
pcDNA3.1-EGFP- SARS-CoV-2 D614G S	This paper	N/A
pET28a-A1L35HR2m	Genewiz	N/A
pET28a-HR2mL35A1	Genewiz	N/A
pET28a-A1L5HR2m	Genewiz	N/A
pET28a-A1HR2m	Genewiz	N/A
pET28a-A3L35HR2m	This paper	N/A
pET28a-A7L35HR2m	This paper	N/A
pET28a-A1L35HR2mdCdN	This paper	N/A
pNL4-3.Luc.RE	Xia et al., 2020²⁰	N/A

Software and algorithms

GraphPad Prism Version 6.02	GraphPad	https://www.graphpad.com
ImageJ	National Institutes of Health	https://imagej.nih.gov/ij

Other

Ni-NTA beads	Smart-Lifesciences	Cat#: SA004100
Amicon column	Millipore	Cat#: UFC5003BK
TissueLyser II	Qiagen	https://www.qiagen.com/au/listpages/ez1-cards/tissuelyser-ii/
SAPPHIRE FL BIOMOLECULAR IMAGER	AZURE Biosystems	Cat#: IS4000
Stainless Steel Beads, 5mm	Qiagen	Cat#: 69989