New guide RNA or fragment comprising its nucleotide sequence, used in composition, kit, Clustered regularly interspaced short palindromic repeats-Cas-based nucleic acid detection system for detecting single nucleotide polymorphism, sickle cell disease, preferably sickle cell anemia
	SARDA A; ARORA S; CHANDRU V; HEGDE V; NAIK M; GUPTA V; ARORA R; SIJI A
	2024-11-05
专利权人	CRISPRBITS PRIVATE LTD (CRIS-Non-standard)
申请日期	2024-11-05
专利号	IN202411084372-A
成果简介	NOVELTY - Guide RNA or a fragment is new, where guide RNA or fragment, comprises nucleotide sequence selected from sequences comprising 20 nucleobases of (SEQ ID NOs: 1 and 2), given in the specification or a sequence having 85% identity with SEQ ID NO: 1 or 2. USE - Guide RNA or fragment used in composition, kit, and clustered regularly interspaced short palindromic repeats (CRISPR)-Cas-based nucleic acid detection system specific for rs334 SNP, Cas12 nuclease, and Cas12b nuclease for detecting single nucleotide polymorphism (SNP), sickle cell disease, preferably sickle cell anemia, and determining homozygous or heterozygous or a carrier state of sickle cell disease (all claimed). ADVANTAGE - The guide RNA recognizes target nucleic acid comprising SNP and directs the Cas12b nuclease to the site of editing. The method determines homozygous or heterozygous or a carrier state of sickle cell disease. The CRISPR-based assay targets specific nucleotide changes with high precision, addressing effectively the variability and specificity issues faced by conventional methods. The method uses specific guide RNA for identifying the specific SNP, that is capable of precisely detecting carrier genotypes. DETAILED DESCRIPTION - Guide RNA or a fragment is new, where guide RNA or fragment, comprises nucleotide sequence selected from sequences comprising 20 nucleobases 5'-tcctcaggagtcagctgcac-3', and 5'-tccacaggagtcagctgcac-3' of (SEQ ID NOs: 1 and 2), or a sequence having 85% identity with SEQ ID NO: 1 or 2. INDEPENDENT CLAIMS are included for: Composition comprising a guide RNA or a fragment and Cas12 nuclease; Kit, which comprises a guide RNA or a fragment, primer or a fragment, Cas 12 nuclease, fluorescent agent or a lateral flow strip along with an instruction manual; Clustered regularly interspaced short palindromic repeats (CRISPR)-Cas-based nucleic acid detection system, which comprises the guide RNA and Cas12 nuclease; and Method for detecting a target nucleic acid or a genotype variant in a biological sample, which involves: (1) preparing a CRISPR reaction mixture using the kit or the guide RNA for genotyping Hemoglobin Subunit Beta (HBB) gene in a biological sample; (2) contacting the biological sample with the CRISPR reaction mixture; and (3) detecting presence of the target nucleic acid or a genotype variant and/or level of the detectable signal.
IPC 分类号	A61K-035/28 ; C07K-014/805 ; C12N-015/11 ; C12N-015/90 ; C12N-009/22
国家	印度
专业领域	医药卫生
语种	英语
成果类型	专利
文献类型	科技成果
条目标识符	http://119.78.100.226:8889/handle/3KE4DYBR/14643
专题	中国科学院新疆生态与地理研究所
作者单位	CRISPRBITS PRIVATE LTD (CRIS-Non-standard)
推荐引用方式 GB/T 7714	SARDA A,ARORA S,CHANDRU V,et al. New guide RNA or fragment comprising its nucleotide sequence, used in composition, kit, Clustered regularly interspaced short palindromic repeats-Cas-based nucleic acid detection system for detecting single nucleotide polymorphism, sickle cell disease, preferably sickle cell anemia. IN202411084372-A[P]. 2024.

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