I designed some asRNA complementary to ssRNA of Severe Acute Respiratory Syndrome CoronaVirus 2 (SARS-CoV-2) and SARS coronavirus 1 (SARS-CoV-1) to limit virus replication through antisense mechanism.
Hybridization with the virus ssRNA can lead to inhibit the specific protein production through translation arrest or degradation by RNase H cleavage.
Has anyone the possibility to check following antisense RNA (asRNA)?
MN988669.1:13467R20_asRNA (gene="orf1ab",ribosomal_slippage)
5' GCA CUU ACA CCG CAA ACC CG 3'
Tm = 91.2°[%GC method]
more antisense RNA
MN988669.1:16842R23_asRNA (gene="orf1ab",ribosomal_slippage)
5' ACA ACA GCA UCA CCA UAG UCA CC 3'
Tm = 85.9°[%GC method]
MN988669.1:28525R21_asRNA (gene="N", structural protein)
5' GGU AGC UCU UCG GUA GUA GCC 3'
Tm = 90.4°[%GC method]
MN988669.1:29013R21_asRNA (gene="N", structural protein)
5' GCC UCA GCA GCA GAU UUC UUA 3'
Tm = 83.7°[%GC method]
MN988669.1:26449R21_asRNA (gene="E", structural protein)
5' AGA CCA GAA GAU CAG GAA CUC 3'
Tm = 83.7°[%GC method]
MN988669.1:29443R21_asRNA (gene="N", structural protein) mismatche in 5' to SARS-CoV-1
5' AGC AGG AAG AAG AGU CAC AGU 3'
Tm = 83.7°[%GC method]
MN988669.1:28789R21_asRNA (gene="N", structural protein) 2 mismatches to SARS-CoV-1
5' GCC GCC UCU GCU CCC UUC UGC 3'
Tm = 100.4°[%GC method]
MN988669.1:27797R21_asRNA (non coding region?)
5' AAC AAG GAA UAG CAG AAA GGC 3'
Tm = 80.4°[%GC method]
Your comments suggestions would be greatly appreciated.
Best Regards
There are many methods for the in vitro transfection like
the classical chemical transfection with calcium phosphate or lipofection.
Other classical examples of physical transfection are electroporation or microinjection.
It's better if you have an etablishied transfection method in your high-security lab.
It's a good idea for detection but for traitment, I have some doubts. How to delivry the antisens RNA in the human body ?
More antisense RNA (asRNA) in highly conserved regions of coronavirus (SARS and MERS):
MN988669.1:14755R24 (gene="orf1ab", ribosomal_slippage,putative RNA-dependent RNA polymerase)
5' UAC CAU CCU GAG CAA AGA AGA AGU 3'
Td = 70.2°[nearest neighbor method]
Tm = 82.5°[%GC method]
Alignment
5' ACTTCTTCTTTGCTCAGGATGGcA 3' SARS-CoV-1
5' ACTTCTTCTTTGCTCAGGATGGTA 3' SARS-CoV-2
5' AtTTtTTCTTTGCTCAaGATGGTA 3' MERS-CoV
MN988669.1:17826R23 (nsp13, putative zinc-binding domain, putative superfamily 1 helicase)
5' CCC UGU GAU GAA UCA ACA GUU UG 3'
Td = 66.7°[nearest neighbor method]
Tm = 82.9°[%GC method]
Alignment
5' CAgACTGTTGATTCATCACAGGG 3' SARS-CoV-1
5' CAAACTGTTGATTCATCACAGGG 3' SARS-CoV-2
5' CAgACTGTTGATTCcTCACAGGG 3' MERS-CoV
PS: Do not expect a finished product.
Laboratories with the appropriate equipment and methods are of course addressed for the check.
Research and Development on Therapeutic Agents and Vaccines for COVID-19 and Related Human Coronavirus Diseases.
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