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
Based on the crystal structure of LbCas12a-crRNA-dsDNA (PDB ID: 5XUS)12, we reasoned that crRNA extensions can influence the trans-cleavage activity by either activating or inhibiting the catalytic efficiency of Cas12a, allowing us to better understand crRNA design with tunable trans-cleavage act...
we optimize the Cas12a system and demonstrate crRNA design principles for this optimized system. Together, these modifications achieve sufficient gene editing efficiency in mammalian cells for Cas12a-based dropout genetic screening. Subsequently, we apply this optimized Cas12a system in a double knockout...
Because both aTFs and the Cas12a–crRNA complex can bind to the same target dsDNA, the presence of both aTF and Cas12a in the same buffer system allows them to compete for and interfere with the same target dsDNA. Therefore, we engineered the aTFs so that they could be immobilized to avo...
The loop primers are provided with the four core primers by the PrimerExplorer design software. Furthermore, two other primer sets that may be added include the “stem primers”, which target the single-stranded region between F1/F2/F3 and B1/B2/B3 (Fig. 3a)52, and the “swarm primer...
摘要 作为世界上最重要的粮食作物之一,水稻生产关系到我国乃至世界的粮食安全。随着人口增 长、全球气候变暖、耕地面积和水资源减少,我国水稻生产面临着巨大挑战。以CRISPR/Cas 系 统为代表的基因编辑技术可对靶标基因进行定点敲除、插入、替换等。大多数农作物优异等位基 因差异主要是少数几个碱基或小片段插入/缺失引...
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...
For SpCas9-NG, these extended coordinates were used to design 20-base-long crRNAs on both the forward and reverse strand using a custom R script utilizing the following PAM sites: AG, TG, CG, GG. In addition, using these extended coordinates, 23-base-long crRNAs for use with (en)As...