Cas12a属于第2大类V型CRISPR/Cas核酸酶,与广泛使用的Cas9(第2大类II型)核酸酶不同,它作为基因组编辑工具有以下几个特性。(1) Cas12a核酸酶与单一CRISPR RNA (crRNA)结合,不需要反式激活CRISPR RNA (tracrRNA),故而Cas12a的crRNA ...
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crRNA Design-Cas12a(Cpf1) Input Sequence General Cpf1 Setting Primers Options Input an DNA sequence in FASTA format OR upload a fasta file: Choose File no file selected Find targets
it is processed into mature crRNAs before itcan form an RNP with the endonuclease. In the case of Cas9, the tracrRNA(encoded close to the CRISPR locus) first needs to hybridize with thepre-crRNA, and then this hybrid RNA–RNA duplex structure is recognized by Cas9,following which the host...
Here, we develop transgene-free canker-resistant Citrus sinensis lines in the T0 generation within 10 months through transformation of embryogenic protoplasts with Cas12a/crRNA ribonucleoprotein to edit the canker susceptibility gene CsLOB1. Among the 39 regenerated lines, 38 are biallelic/homozygous ...
Compared with Cas9, Cas12 requires a shorter CRISPR RNA (crRNA) and has an additional trans-cleavage activity to cleave single-stranded DNA with an arbitrary sequence, which has led to Cas12 being more widely used in the field of genetic engineering in recent years. Combining CRISPR/Cas with...
Here, we design and validate sixteen types of structurally engineered Cas12a crRNAs targeting various immunologically relevant loci in-vitro and in-cellulo. We show that all our structural modifications in the loop region, ranging from engineered breaks (STAR-crRNAs) to large gaps (Gap-crRNAs),...
It has been proven that the Cas12a protein assembles with CRISPR RNA (crRNA) to form a Cas12a/crRNA complex, and the indiscriminate (nonspecific) single-stranded DNase activity of CRISPR/Cas12a can be activated by both single- and double-stranded target DNA [15], [16]. Compared with ...
具体实施方式 [0139] 本发明人经过广泛而深入地研究,经过大量的筛选,首次意外地发现了一批特异 性crRNA,结合CRISPR/Cas12a检测体系,建立了对GBS的快速检测体系及方法,包括荧光检 测方法及胶体金检测方法,极大地缩短了对GBS的检测时间。在此基础上完成了本发明。 9 9 CN 116179723 A 说明书 7/16页 [0140] 术语...
野生型AsCpf1与以下比较:(A)T167/T539变体,使用靶向EGFP中的位点的crRNA;(B)S170和E174变体,使用靶向EGFP中的位点的crRNA;(C和D)S542变体,使用靶向EGFP中的位点(子图C)或内源人类基因位点(子图D)的crRNA;(E)N551和N552变体,使用靶向EGFP中的位点的crRNA;(F)K607变体,使用靶向EGFP中的位点的crRNA;(G和...