CN114250229B - 抑制新冠病毒2019-nCoV的siRNA及其用途 - Google Patents
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Abstract
本发明提供的一种抑制新冠病毒2019‑nCoV的siRNA,所述siRNA为如下双链RNA序列中的至少一种:2019‑nCoV‑siRNA1‑6:核苷酸序列如SEQ ID NO:1‑12所示;本发明发现选择新冠病毒2019‑nCoV(登记号NC_045512)的保守区S基因中位点cd21591附近能够设计得到高效抑制新冠病毒2019‑nCoV的siRNA,对于临床抗新冠病毒2019‑nCoV药物的开发具有重要意义。
Description
技术领域
本发明涉及RNA干扰抗病毒领域,具体涉及一种抑制新冠病毒 2019-nCoV的siRNA及其用途。
背景技术
冠状病毒是一个大型病毒家族,已知可引起感冒以及中东呼吸综合征 (MERS)和严重急性呼吸综合征(SARS)等较严重疾病。
人感染了冠状病毒后常见体征有呼吸道症状、发热、咳嗽、气促和呼吸困难等。在较严重病例中,感染可导致肺炎、严重急性呼吸综合征、肾衰竭,甚至死亡。2020年8月,最新研究成果揭示新冠病毒的传播特征:高传染性和高隐蔽性。
RNA干扰(RNA interference,RNAi)是指在进化过程中高度保守的、由双链RNA(double-stranded RNA,dsRNA)诱发的、同源mRNA高效特异性降解的现象。基因沉默,主要有转录前水平的基因沉默(TGS)和转录后水平的基因沉默(PTGS)两类:TGS是指由于DNA修饰或染色体异染色质化等原因使基因不能正常转录;PTGS是启动了细胞质内靶mRNA序列特异性的降解机制。有时转基因会同时导致TGS和PTGS。由于使用RNAi技术可以特异性剔除或关闭特定基因的表达,(长度超过三十的dsRNA会引起干扰素毒性)所以该技术已被广泛用于探索基因功能和传染性疾病及恶性肿瘤的治疗领域。因此,本发明提出了一种抑制新冠病毒2019-nCoV的 siRNA及其用途。
发明内容
因此,本发明要解决的技术问题在于提供一种抑制新冠病毒2019-nCoV 的siRNA及其用途。
本发明提供了一种抑制新冠病毒2019-nCoV的siRNA,所述siRNA为如下双链RNA序列中的至少一种:
2019-nCoV-siRNA1:正义链如SEQ ID NO:1所示,反义链如SEQ ID NO:2所示;
2019-nCoV-siRNA2:正义链如SEQ ID NO:3所示,反义链如SEQ ID NO:4所示;
2019-nCoV-siRNA3:正义链如SEQ ID NO:5所示,反义链如SEQ ID NO:6所示;
2019-nCoV-siRNA4:正义链如SEQ ID NO:7所示,反义链如SEQ ID NO:8所示;
2019-nCoV-siRNA5:正义链如SEQ ID NO:9所示,反义链如SEQ ID NO:10所示;
2019-nCoV-siRNA6:正义链如SEQ ID NO:11所示,反义链如SEQ ID NO:12所示。
所述的抑制新冠病毒2019-nCoV的siRNA,所述siRNA为如下双链 RNA序列中的至少一种:
1)与2019-nCoV-siRNA1的双链RNA序列具有85%以上的同源性,且具有抑制冠病毒2019-nCoV的作用;
2)与2019-nCoV-siRNA2的双链RNA序列具有85%以上的同源性,且具有抑制冠病毒2019-nCoV的作用;
3)与2019-nCoV-siRNA3的双链RNA序列具有85%以上的同源性,且具有抑制冠病毒2019-nCoV的作用;
4)与2019-nCoV-siRNA4的双链RNA序列具有85%以上的同源性,且具有抑制冠病毒2019-nCoV的作用;
5)与2019-nCoV-siRNA5的双链RNA序列具有85%以上的同源性,且具有抑制冠病毒2019-nCoV的作用;
6)与2019-nCoV-siRNA6的双链RNA序列具有85%以上的同源性,且具有抑制冠病毒2019-nCoV的作用。
所述的抑制新冠病毒2019-nCoV的siRNA,所述siRNA为双链RNA 序列中的至少一条单链的5’末端进行磷酸化修饰。
所述的抑制新冠病毒2019-nCoV的siRNA,所述siRNA为双链RNA 序列中正义链的5’末端进行磷酸化修饰。
所述的抑制新冠病毒2019-nCoV的siRNA,所述siRNA为双链RNA 序列中的至少一条单链的5’末端经过胆固醇修饰。
所述的抑制新冠病毒2019-nCoV的siRNA在制备抑制新冠病毒 2019-nCoV的产品中的用途。
所述的用途,所述的产品可以为试剂盒、制剂或药物。
本发明提供了一种抑制新冠病毒2019-nCoV的试剂盒,包括所述的抑制新冠病毒2019-nCoV的siRNA。
本发明技术方案,具有如下优点:
1.本发明提供的一种抑制新冠病毒2019-nCoV的siRNA,所述siRNA 为如下双链RNA序列中的至少一种:2019-nCoV-siRNA1:正义链如SEQ ID NO:1所示,反义链如SEQ IDNO:2所示;2019-nCoV-siRNA2:正义链如SEQ ID NO:3所示,反义链如SEQ ID NO:4所示;2019-nCoV-siRNA3:正义链如SEQ ID NO:5所示,反义链如SEQ ID NO:6所示;2019-nCoV-siRNA4:正义链如SEQ ID NO:7所示,反义链如SEQ ID NO: 8所示;2019-nCoV-siRNA5:正义链如SEQ ID NO:9所示,反义链如SEQ ID NO:10所示;2019-nCoV-siRNA6:正义链如SEQ IDNO:11所示,反义链如SEQ ID NO:12所示;本发明发现选择新冠病毒2019-nCoV(登记号NC_045512)的保守区S基因中位点cd21591附近能够设计得到高效抑制新冠病毒2019-nCoV的siRNA,对于临床抗新冠病毒2019-nCoV药物的开发具有重要意义。
附图说明
为了更清楚地说明本发明具体实施方式或现有技术中的技术方案,下面将对具体实施方式或现有技术描述中所需要使用的附图作简单地介绍,显而易见地,下面描述中的附图是本发明的一些实施方式,对于本领域普通技术人员来讲,在不付出创造性劳动的前提下,还可以根据这些附图获得其他的附图。
图1是本发明实验例1中质粒pcDNA6.2-GW/EmGFP-miR图谱。
具体实施方式
提供下述实施例是为了更好地进一步理解本发明,并不局限于所述最佳实施方式,不对本发明的内容和保护范围构成限制,任何人在本发明的启示下或是将本发明与其他现有技术的特征进行组合而得出的任何与本发明相同或相近似的产品,均落在本发明的保护范围之内。
实施例中未注明具体实验步骤或条件者,按照本领域内的文献所描述的常规实验步骤的操作或条件即可进行。所用试剂或仪器未注明生产厂商者,均为可以通过市购获得的常规试剂产品。
实施例1抑制新冠病毒2019-nCoV的siRNA的合成和修饰
本发明发现选择新冠病毒2019-nCoV(登记号NC_045512)的S基因 (如SEQ ID NO:87所示,登记号:43740568)cd21591位点附近,能够设计得到高效抑制新冠病毒2019-nCoV的siRNA(如下表1)。设计的siRNA 由上海权阳生物科技有限公司合成,在合成时,同时在所述siRNA为双链 RNA序列中正义链的5’末端进行磷酸化修饰。
表1
实验例1
本实验考查了设计合成的siRNA对新冠病毒2019-nCoV的抑制作用,考察的siRNA如下表2所示,表2中的siRNA均以新冠病毒2019-nCoV(登记号NC_045512)的S基因(如SEQID NO:87所示,登记号:43740568) 为靶序列,参考siRNA设计原则设计的,所述siRNA为双链RNA序列中正义链的5’末端进行磷酸化修饰,正义链的3’末端进行胆固醇修饰。
表2siRNA
(1)质粒合成
将新冠病毒2019-nCoV的保守区基因序列(由上海生工生物工程公司合成和测序)插入质粒pcDNA6.2-GW/EmGFP-miR(图谱如图1所示,为市售产品)中,插入序列为SEQ IDNO:87,BamHI和BglII,制备得到质粒pcDNA6.2-GW/EmGFP-siRNA(由上海生工生物工程公司合成和测序)。
(2)转染
将9ulRNAiMAX Reagent用150ul无血清opti-MEM 稀释,并将30pmol(加入的siRNA总浓度,加入的siRNA参见表3,其中 2019-nCoV-siRNA1+2019-nCoV-siRNA2表示两siRNA等摩尔比混合的组合 siRNA,同理2019-nCoV-siRNA3+2019-nCoV-siRNA5表示两siRNA等摩尔比混合的组合siRNA,2019-nCoV-siRNA4+2019-nCoV-siRNA6表示两 siRNA等摩尔比混合的组合siRNA,空白对照组加入等体积的蒸馏水) siRNA用150ul无血清opti-MEM稀释,两份稀释液混合,室温孵育5min 后转入60-70%融合度的呼吸道上皮细胞系的HBE细胞中进行转染。
(3)实时荧光定量PCR检测siRNA对新冠病毒2019-nCoV的抑制效果
1)、提取步骤(2)中的转染后的细胞的总RNA,提取试剂选择天根 RNA Easy Fast动物组织/细胞总RNA提取试剂盒(DP451),然后将提取的总RNA逆转录成cDNA,逆转录试剂盒选择TAKARA反转录试剂盒PrimeScript RT Master Mix;
提取总RNA:收集细胞离心,加入适量裂解液RLA和10μl Proteinase K,旋涡震荡;离心取上清加入基因组DNA去除柱中,吸附和清洗后用洗脱液洗脱收集。逆转录:按照试剂盒提供配方配制反转录体系,轻柔混匀后进行逆转录反应。
2)、采用实时荧光定量PCR试剂盒TAKARA实时荧光定量试剂盒观察对siRNA对新冠病毒2019-nCoV的抑制效果。
按照试剂盒中组分配制PCR反应液,依据两步法PCR反应程序进行实时荧光定量PCR反应。
抑制率效果如下表3:
表3抑制率效果
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由以上结果可知,本发明在S基因(如SEQ ID NO:87所示,登记号:43740568)cd21591位点附近设计的siRNA在抑制新冠病毒2019-nCoV 方面具有高效率,对于临床抗新冠病毒2019-nCoV药物的开发具有重要意义,而当将2019-nCoV-siRNA1和2019-nCoV-siRNA2组合使用时,具有一定的协同作用,能够显著提高对新冠病毒2019-nCoV的抑制作用。
显然,上述实施例仅仅是为清楚地说明所作的举例,而并非对实施方式的限定。对于所属领域的普通技术人员来说,在上述说明的基础上还可以做出其它不同形式的变化或变动。这里无需也无法对所有的实施方式予以穷举。而由此所引伸出的显而易见的变化或变动仍处于本发明创造的保护范围之中。
序列表
<110> 深圳大学
深圳大学总医院
<120> 抑制新冠病毒2019-nCoV的siRNA及其用途
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<213> 人工序列(Artificial Sequence)
<400> 11
gguacuaaga gguuugauaa c 21
<210> 12
<211> 21
<212> RNA
<213> 人工序列(Artificial Sequence)
<400> 12
uaucaaaccu cuuaguacca u 21
<210> 13
<211> 21
<212> RNA
<213> 人工序列(Artificial Sequence)
<400> 13
guacuacuuu agauucgaag a 21
<210> 14
<211> 21
<212> RNA
<213> 人工序列(Artificial Sequence)
<400> 14
uucgaaucua aaguaguacc a 21
<210> 15
<211> 21
<212> RNA
<213> 人工序列(Artificial Sequence)
<400> 15
gaauuugugu uuaagaauau u 21
<210> 16
<211> 21
<212> RNA
<213> 人工序列(Artificial Sequence)
<400> 16
uauucuuaaa cacaaauucc c 21
<210> 17
<211> 21
<212> RNA
<213> 人工序列(Artificial Sequence)
<400> 17
uauucuuaaa cacaaauucc c 21
<210> 18
<211> 21
<212> RNA
<213> 人工序列(Artificial Sequence)
<400> 18
uaacaauaag uagggacugg g 21
<210> 19
<211> 21
<212> RNA
<213> 人工序列(Artificial Sequence)
<400> 19
ggaauuugug uuuaagaaua u 21
<210> 20
<211> 21
<212> RNA
<213> 人工序列(Artificial Sequence)
<400> 20
auucuuaaac acaaauuccc u 21
<210> 21
<211> 21
<212> RNA
<213> 人工序列(Artificial Sequence)
<400> 21
ugagaagucu aacauaauaa g 21
<210> 22
<211> 21
<212> RNA
<213> 人工序列(Artificial Sequence)
<400> 22
uauuauguua gacuucucag u 21
<210> 23
<211> 21
<212> RNA
<213> 人工序列(Artificial Sequence)
<400> 23
gaaagugagu ucagaguuua u 21
<210> 24
<211> 21
<212> RNA
<213> 人工序列(Artificial Sequence)
<400> 24
aaacucugaa cucacuuucc a 21
<210> 25
<211> 21
<212> RNA
<213> 人工序列(Artificial Sequence)
<400> 25
guugaaaucc uucacuguag a 21
<210> 26
<211> 21
<212> RNA
<213> 人工序列(Artificial Sequence)
<400> 26
uacagugaag gauuucaacg u 21
<210> 27
<211> 21
<212> RNA
<213> 人工序列(Artificial Sequence)
<400> 27
aguuuauucu agugcgaaua a 21
<210> 28
<211> 21
<212> RNA
<213> 人工序列(Artificial Sequence)
<400> 28
auucgcacua gaauaaacuc u 21
<210> 29
<211> 21
<212> RNA
<213> 人工序列(Artificial Sequence)
<400> 29
gaguucagag uuuauucuag u 21
<210> 30
<211> 21
<212> RNA
<213> 人工序列(Artificial Sequence)
<400> 30
uagaauaaac ucugaacuca c 21
<210> 31
<211> 21
<212> RNA
<213> 人工序列(Artificial Sequence)
<400> 31
agauuuccua auauuacaaa c 21
<210> 32
<211> 21
<212> RNA
<213> 人工序列(Artificial Sequence)
<400> 32
uuguaauauu aggaaaucua a 21
<210> 33
<211> 21
<212> RNA
<213> 人工序列(Artificial Sequence)
<400> 33
uacuaauguu guuauuaaag u 21
<210> 34
<211> 21
<212> RNA
<213> 人工序列(Artificial Sequence)
<400> 34
uuuaauaaca acauuaguag c 21
<210> 35
<211> 21
<212> RNA
<213> 人工序列(Artificial Sequence)
<400> 35
agcuggugcu gcagcuuauu a 21
<210> 36
<211> 21
<212> RNA
<213> 人工序列(Artificial Sequence)
<400> 36
auaagcugca gcaccagcug u 21
<210> 37
<211> 21
<212> RNA
<213> 人工序列(Artificial Sequence)
<400> 37
gaacauguca acaacucaua u 21
<210> 38
<211> 21
<212> RNA
<213> 人工序列(Artificial Sequence)
<400> 38
augaguuguu gacauguuca g 21
<210> 39
<211> 21
<212> RNA
<213> 人工序列(Artificial Sequence)
<400> 39
acacgugcag gcuguuuaau a 21
<210> 40
<211> 21
<212> RNA
<213> 人工序列(Artificial Sequence)
<400> 40
uuaaacagcc ugcacguguu u 21
<210> 41
<211> 21
<212> RNA
<213> 人工序列(Artificial Sequence)
<400> 41
caguguguua aucuuacaac c 21
<210> 42
<211> 21
<212> RNA
<213> 人工序列(Artificial Sequence)
<400> 42
uuguaagauu aacacacuga c 21
<210> 43
<211> 21
<212> RNA
<213> 人工序列(Artificial Sequence)
<400> 43
ccagugucua ugaccaagac a 21
<210> 44
<211> 21
<212> RNA
<213> 人工序列(Artificial Sequence)
<400> 44
ucuuggucau agacacuggu a 21
<210> 45
<211> 21
<212> RNA
<213> 人工序列(Artificial Sequence)
<400> 45
guacuaagag guuugauaac c 21
<210> 46
<211> 21
<212> RNA
<213> 人工序列(Artificial Sequence)
<400> 46
uuaucaaacc ucuuaguacc a 21
<210> 47
<211> 21
<212> RNA
<213> 人工序列(Artificial Sequence)
<400> 47
agauugcuga uuauaauuau a 21
<210> 48
<211> 21
<212> RNA
<213> 人工序列(Artificial Sequence)
<400> 48
uaauuauaau cagcaaucuu u 21
<210> 49
<211> 21
<212> RNA
<213> 人工序列(Artificial Sequence)
<400> 49
agaaucuauu guuagauuuc c 21
<210> 50
<211> 21
<212> RNA
<213> 人工序列(Artificial Sequence)
<400> 50
aaaucuaaca auagauucug u 21
<210> 51
<211> 21
<212> RNA
<213> 人工序列(Artificial Sequence)
<400> 51
acccuguccu accauuuaau g 21
<210> 52
<211> 21
<212> RNA
<213> 人工序列(Artificial Sequence)
<400> 52
uuaaauggua ggacaggguu a 21
<210> 53
<211> 21
<212> RNA
<213> 人工序列(Artificial Sequence)
<400> 53
agguuggugg uaauuauaau u 21
<210> 54
<211> 21
<212> RNA
<213> 人工序列(Artificial Sequence)
<400> 54
uuauaauuac caccaaccuu a 21
<210> 55
<211> 21
<212> RNA
<213> 人工序列(Artificial Sequence)
<400> 55
guggguuauc uucaaccuag g 21
<210> 56
<211> 21
<212> RNA
<213> 人工序列(Artificial Sequence)
<400> 56
uagguugaag auaacccaca u 21
<210> 57
<211> 21
<212> RNA
<213> 人工序列(Artificial Sequence)
<400> 57
caaugguacu aagagguuug a 21
<210> 58
<211> 21
<212> RNA
<213> 人工序列(Artificial Sequence)
<400> 58
aaaccucuua guaccauugg u 21
<210> 59
<211> 21
<212> RNA
<213> 人工序列(Artificial Sequence)
<400> 59
guguucuuac ugagucuaac a 21
<210> 60
<211> 21
<212> RNA
<213> 人工序列(Artificial Sequence)
<400> 60
uuagacucag uaagaacacc u 21
<210> 61
<211> 21
<212> RNA
<213> 人工序列(Artificial Sequence)
<400> 61
aagucuacua auuugguuaa a 21
<210> 62
<211> 21
<212> RNA
<213> 人工序列(Artificial Sequence)
<400> 62
uaaccaaauu aguagacuuu u 21
<210> 63
<211> 21
<212> RNA
<213> 人工序列(Artificial Sequence)
<400> 63
gaggugauga agucagacaa a 21
<210> 64
<211> 21
<212> RNA
<213> 人工序列(Artificial Sequence)
<400> 64
ugucugacuu caucaccucu a 21
<210> 65
<211> 21
<212> RNA
<213> 人工序列(Artificial Sequence)
<400> 65
agguugcugu ucuuuaucag g 21
<210> 66
<211> 21
<212> RNA
<213> 人工序列(Artificial Sequence)
<400> 66
ugauaaagaa cagcaaccug g 21
<210> 67
<211> 21
<212> RNA
<213> 人工序列(Artificial Sequence)
<400> 67
gaaccacaaa ucauuacuac a 21
<210> 68
<211> 21
<212> RNA
<213> 人工序列(Artificial Sequence)
<400> 68
uaguaaugau uugugguuca u 21
<210> 69
<211> 21
<212> RNA
<213> 人工序列(Artificial Sequence)
<400> 69
guuucugccu uuccaacaau u 21
<210> 70
<211> 21
<212> RNA
<213> 人工序列(Artificial Sequence)
<400> 70
uuguuggaaa ggcagaaacu u 21
<210> 71
<211> 21
<212> RNA
<213> 人工序列(Artificial Sequence)
<400> 71
gcaaauuuga ugaagacgac u 21
<210> 72
<211> 21
<212> RNA
<213> 人工序列(Artificial Sequence)
<400> 72
ucgucuucau caaauuugca g 21
<210> 73
<211> 21
<212> RNA
<213> 人工序列(Artificial Sequence)
<400> 73
ccaauuuaau agugcuauug g 21
<210> 74
<211> 21
<212> RNA
<213> 人工序列(Artificial Sequence)
<400> 74
aauagcacua uuaaauuggu u 21
<210> 75
<211> 21
<212> RNA
<213> 人工序列(Artificial Sequence)
<400> 75
acguagugua gcuagucaau c 21
<210> 76
<211> 21
<212> RNA
<213> 人工序列(Artificial Sequence)
<400> 76
uugacuagcu acacuacgug c 21
<210> 77
<211> 21
<212> RNA
<213> 人工序列(Artificial Sequence)
<400> 77
gcuuuaaaca cgcuuguuaa a 21
<210> 78
<211> 21
<212> RNA
<213> 人工序列(Artificial Sequence)
<400> 78
uaacaagcgu guuuaaagcu u 21
<210> 79
<211> 21
<212> RNA
<213> 人工序列(Artificial Sequence)
<400> 79
gugucuauga ccaagacauc a 21
<210> 80
<211> 21
<212> RNA
<213> 人工序列(Artificial Sequence)
<400> 80
augucuuggu cauagacacu g 21
<210> 81
<211> 21
<212> RNA
<213> 人工序列(Artificial Sequence)
<400> 81
gcucaaagga gucaaauuac a 21
<210> 82
<211> 21
<212> RNA
<213> 人工序列(Artificial Sequence)
<400> 82
uaauuugacu ccuuugagca c 21
<210> 83
<211> 21
<212> RNA
<213> 人工序列(Artificial Sequence)
<400> 83
cagaaucuau uguuagauuu c 21
<210> 84
<211> 21
<212> RNA
<213> 人工序列(Artificial Sequence)
<400> 84
aaucuaacaa uagauucugu u 21
<210> 85
<211> 21
<212> DNA/RNA
<213> 人工序列(Artificial Sequence)
<400> 85
uucuccguca cgacgaugut t 21
<210> 86
<211> 21
<212> DNA/RNA
<213> 人工序列(Artificial Sequence)
<400> 86
acgugggcga guuacacaat t 21
<210> 87
<211> 3822
<212> DNA/RNA
<213> 新型冠状病毒(2019-nCoV)(新型冠状病毒(2019-nCoV))
<400> 87
atgtttgttt ttcttgtttt attgccacta gtctctagtc agtgtgttaa tcttacaacc 60
agaactcaat taccccctgc atacactaat tctttcacac gtggtgttta ttaccctgac 120
aaagttttca gatcctcagt tttacattca actcaggact tgttcttacc tttcttttcc 180
aatgttactt ggttccatgc tatacatgtc tctgggacca atggtactaa gaggtttgat 240
aaccctgtcc taccatttaa tgatggtgtt tattttgctt ccactgagaa gtctaacata 300
ataagaggct ggatttttgg tactacttta gattcgaaga cccagtccct acttattgtt 360
aataacgcta ctaatgttgt tattaaagtc tgtgaatttc aattttgtaa tgatccattt 420
ttgggtgttt attaccacaa aaacaacaaa agttggatgg aaagtgagtt cagagtttat 480
tctagtgcga ataattgcac ttttgaatat gtctctcagc cttttcttat ggaccttgaa 540
ggaaaacagg gtaatttcaa aaatcttagg gaatttgtgt ttaagaatat tgatggttat 600
tttaaaatat attctaagca cacgcctatt aatttagtgc gtgatctccc tcagggtttt 660
tcggctttag aaccattggt agatttgcca ataggtatta acatcactag gtttcaaact 720
ttacttgctt tacatagaag ttatttgact cctggtgatt cttcttcagg ttggacagct 780
ggtgctgcag cttattatgt gggttatctt caacctagga cttttctatt aaaatataat 840
gaaaatggaa ccattacaga tgctgtagac tgtgcacttg accctctctc agaaacaaag 900
tgtacgttga aatccttcac tgtagaaaaa ggaatctatc aaacttctaa ctttagagtc 960
caaccaacag aatctattgt tagatttcct aatattacaa acttgtgccc ttttggtgaa 1020
gtttttaacg ccaccagatt tgcatctgtt tatgcttgga acaggaagag aatcagcaac 1080
tgtgttgctg attattctgt cctatataat tccgcatcat tttccacttt taagtgttat 1140
ggagtgtctc ctactaaatt aaatgatctc tgctttacta atgtctatgc agattcattt 1200
gtaattagag gtgatgaagt cagacaaatc gctccagggc aaactggaaa gattgctgat 1260
tataattata aattaccaga tgattttaca ggctgcgtta tagcttggaa ttctaacaat 1320
cttgattcta aggttggtgg taattataat tacctgtata gattgtttag gaagtctaat 1380
ctcaaacctt ttgagagaga tatttcaact gaaatctatc aggccggtag cacaccttgt 1440
aatggtgttg aaggttttaa ttgttacttt cctttacaat catatggttt ccaacccact 1500
aatggtgttg gttaccaacc atacagagta gtagtacttt cttttgaact tctacatgca 1560
ccagcaactg tttgtggacc taaaaagtct actaatttgg ttaaaaacaa atgtgtcaat 1620
ttcaacttca atggtttaac aggcacaggt gttcttactg agtctaacaa aaagtttctg 1680
cctttccaac aatttggcag agacattgct gacactactg atgctgtccg tgatccacag 1740
acacttgaga ttcttgacat tacaccatgt tcttttggtg gtgtcagtgt tataacacca 1800
ggaacaaata cttctaacca ggttgctgtt ctttatcagg atgttaactg cacagaagtc 1860
cctgttgcta ttcatgcaga tcaacttact cctacttggc gtgtttattc tacaggttct 1920
aatgtttttc aaacacgtgc aggctgttta ataggggctg aacatgtcaa caactcatat 1980
gagtgtgaca tacccattgg tgcaggtata tgcgctagtt atcagactca gactaattct 2040
cctcggcggg cacgtagtgt agctagtcaa tccatcattg cctacactat gtcacttggt 2100
gcagaaaatt cagttgctta ctctaataac tctattgcca tacccacaaa ttttactatt 2160
agtgttacca cagaaattct accagtgtct atgaccaaga catcagtaga ttgtacaatg 2220
tacatttgtg gtgattcaac tgaatgcagc aatcttttgt tgcaatatgg cagtttttgt 2280
acacaattaa accgtgcttt aactggaata gctgttgaac aagacaaaaa cacccaagaa 2340
gtttttgcac aagtcaaaca aatttacaaa acaccaccaa ttaaagattt tggtggtttt 2400
aatttttcac aaatattacc agatccatca aaaccaagca agaggtcatt tattgaagat 2460
ctacttttca acaaagtgac acttgcagat gctggcttca tcaaacaata tggtgattgc 2520
cttggtgata ttgctgctag agacctcatt tgtgcacaaa agtttaacgg ccttactgtt 2580
ttgccacctt tgctcacaga tgaaatgatt gctcaataca cttctgcact gttagcgggt 2640
acaatcactt ctggttggac ctttggtgca ggtgctgcat tacaaatacc atttgctatg 2700
caaatggctt ataggtttaa tggtattgga gttacacaga atgttctcta tgagaaccaa 2760
aaattgattg ccaaccaatt taatagtgct attggcaaaa ttcaagactc actttcttcc 2820
acagcaagtg cacttggaaa acttcaagat gtggtcaacc aaaatgcaca agctttaaac 2880
acgcttgtta aacaacttag ctccaatttt ggtgcaattt caagtgtttt aaatgatatc 2940
ctttcacgtc ttgacaaagt tgaggctgaa gtgcaaattg ataggttgat cacaggcaga 3000
cttcaaagtt tgcagacata tgtgactcaa caattaatta gagctgcaga aatcagagct 3060
tctgctaatc ttgctgctac taaaatgtca gagtgtgtac ttggacaatc aaaaagagtt 3120
gatttttgtg gaaagggcta tcatcttatg tccttccctc agtcagcacc tcatggtgta 3180
gtcttcttgc atgtgactta tgtccctgca caagaaaaga acttcacaac tgctcctgcc 3240
atttgtcatg atggaaaagc acactttcct cgtgaaggtg tctttgtttc aaatggcaca 3300
cactggtttg taacacaaag gaatttttat gaaccacaaa tcattactac agacaacaca 3360
tttgtgtctg gtaactgtga tgttgtaata ggaattgtca acaacacagt ttatgatcct 3420
ttgcaacctg aattagactc attcaaggag gagttagata aatattttaa gaatcataca 3480
tcaccagatg ttgatttagg tgacatctct ggcattaatg cttcagttgt aaacattcaa 3540
aaagaaattg accgcctcaa tgaggttgcc aagaatttaa atgaatctct catcgatctc 3600
caagaacttg gaaagtatga gcagtatata aaatggccat ggtacatttg gctaggtttt 3660
atagctggct tgattgccat agtaatggtg acaattatgc tttgctgtat gaccagttgc 3720
tgtagttgtc tcaagggctg ttgttcttgt ggatcctgct gcaaatttga tgaagacgac 3780
tctgagccag tgctcaaagg agtcaaatta cattacacat aa 3822
Claims (2)
1.一种抑制新冠病毒2019-nCoV的siRNA在制备抑制新冠病毒2019-nCoV的产品中的用途,其特征在于,所述siRNA为如下2019-nCoV-siRNA1~2019-nCoV-siRNA6中的至少一种:
2019-nCoV-siRNA1:正义链如SEQ ID NO:1所示,反义链如SEQ ID NO:2所示;
2019-nCoV-siRNA2:正义链如SEQ ID NO:3所示,反义链如SEQ ID NO:4所示;
2019-nCoV-siRNA3:正义链如SEQ ID NO:5所示,反义链如SEQ ID NO:6所示;
2019-nCoV-siRNA4:正义链如SEQ ID NO:7所示,反义链如SEQ ID NO:8所示;
2019-nCoV-siRNA5:正义链如SEQ ID NO:9所示,反义链如SEQ ID NO:10所示;
2019-nCoV-siRNA6:正义链如SEQ ID NO:11所示,反义链如SEQ ID NO:12所示;
所述siRNA为双链RNA序列中正义链的5’末端进行磷酸化修饰,正义链的3’末端进行胆固醇修饰。
2.根据权利要求1所述的用途,其特征在于,所述的产品为试剂盒或药物。
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