会出来很多可以用的sgRNA,根据自己的需要进行选择,一般找评分靠前的位置。 备注:还有很多其它的网站,都可以使用,如:CRISPR RGEN Tools (rgenome.net);CRISPOR (tefor.net)。网站时不时会崩一下,这几个网站可以替代使用。谨慎的同学也可以联合几个网站,挑选出最合适的sgRNA。每个在线网站都有评分的侧重点,感兴趣的...
CRISPR gRNA 设计网页汇总 (百度一下以下关键词,第一个结果就是) #1 CRISPR Guide RNA Design Software for Molecular Biology | Benchling #2 sgRNA Designer: CRISPRko (broadinstitute.org) #3 CRISPR RGEN Tools (rgenome.net) #4 CHOPCHOP (uib.no) #5 CRISPOR (tefor.net) #6 E-CRISP Design (e-...
Raw reads of paired-end Miseq sequencing were joined by the programme ‘fastq-join’, and indel frequency and patterns in joined reads were analysed using the Cas-Analyzer programme implemented in CRISPR RGEN Tools (http://rgenome.ibs.re.kr). Confocal laser scanning microscopy The Cy3-...
在仅Agro转化和FrCas9植物中,鉴定出的单核苷酸变异(SNVs)数量相似,然而表达TREX2-FrCas9的植物在C:G>T:A和A:T>T:A突变的数量上有显著增加,表明TREX2-FrCas9会导致更多的核苷酸替换突变,尤其是C:G>T:A和A:T>T:A类型的非目标突变。 此外,研究者利用CRISPR RGEN Tools在线工具来预测依赖于sgRNA的潜在脱靶...
首先利用crispr rgen tools,以靶标(seq id no:1)为模板,设定参数:grna长度为20bp、score分值大于66,选取满足以上参数的8条grna序列作为候选grna序列,详见seq id no:2-9,这8条grna靶向保守区内不同位置的不同的pam序列。依据荧光强度筛选出最优grna序列后,依据其结合位点所在区域再设计筛选出最优rpa引物。
The base editing efficiencies were analysed by the CRISPR RGEN tools. Extended Data Fig. 7 Undetectable off-target genome-editing activity by Cas9-Act3.0 and CBE-Cas9n-Act3.0 at the AtFT promoter with 15-nt protospacers. a, Identification of Cas9-Act3.0-mediated potential indel mutations at ...
首先利用crisprrgen tools,以靶标(seq id no:1)为模板,设定参数:grna长度为20bp、score分值大于66,选取满足以上参数的2条grna序列作为候选grna序列,这2条grna靶向保守区内不同位置的不同的pam序列。依据荧光强度筛选出最优grna序列后,依据其结合位点所在区域再设计筛选出最优rpa引物。
在线设计sgRNA是常用方法，如张峰老师实验室的网站、CRISPR RGEN Tools、CRISPOR等，提供多种设计选项。选择评分靠前的位置进行使用。这些在线工具各有侧重，用户可根据需求选择最适合自己的。对于特定位点编辑的实验，如Knock-in实验，可能需要直接在软件如Snapgene中设计sgRNA。在编码区寻找NGG序列及其前方20个...
CRISPR RGEN Tools is a CRISIPR/Cas library platform that contains multiple sgRNA design tools. For example, CRISPR RGEN Tools employs Cas-designer for conventional CRISPR/Cas nucleases, BE-Designer for CRISPR base editing, and PE-Designer for CRISPR prime editing [93]. In addition, PE-Designer...
Results were analyzed with CRISPR RGEN Tools Cas-Analyser software60. Animal experiments All experiments with mice conformed to Animal Welfare guidelines and were performed in accordance to protocols approved by the Ethics Committee of the Instituto de Salud Carlos III. Tumors were generated in 8-...