This invention relates to recombinant Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR) arrays and recombinant nucleic acid constructs encoding Type l-E CASCADE complexes as well as plasmids, retroviruses and bacteriophage comprising the same. Specifically, the CRISPR array comprising two or...
Class 2 CRISPR–Cas systems, such as Cas9 and Cas12, have been widely used to target DNA sequences in eukaryotic genomes. However, class 1 CRISPR–Cas systems, which represent about 90% of all CRISPR systems in nature, remain largely unexplored for genome engineering applications. Here, we ...
Figure 4. Genome editing in C. pasteurianum using the endogenous Type I-B CRISPR-Cas system. 图4(a)使用内源Type I-B CRISPR-Cas机制的cpaAIR基因删除策略。显示了精简的巴氏梭菌Type I-B CRISPR阵列(阵列)和cas基因操纵子(cas),以及cpaAIR靶向位点。提供了一个插图,显示了由37个间隔阵列和cas操纵子组...
Class 2 CRISPR–Cas proteins have been widely developed as genome editing and transcriptional regulating tools. Class 1 type I CRISPR–Cas constitutes ~60% of all the CRISPR–Cas systems. However, only type I–B and I–E systems have been used to control mammalian gene expression and for gen...
Fig. 1: The crystal structure of AcrIF24, a dimer that inhibits the type I-F CRISPR-Cas system. a AcrIF24 but not AcrIF24ΔMD inhibits the in vitro cleavage activity of the type I-F CRISPR-Cas system. Reactions were performed with 0.32 μM Csy complex, 0.16 μM Cas2/3,...
近日,中山大学生命科学院的科学家团队首次把type I-F CRSIPR-Cas系统Cascade复合物的Csy3亚基与VPR(VP64-p65-Rta)融合, 可以高效激活哺乳动物细胞内源基因的表达。 该研究成果发表在Nature Communications上,题目为Repurposing Type I-F CRISPR-Cas System as a Transcriptional Activation Tool in Human Cells。
对病原DNA的免疫识别，是宿主细胞感知病原感染的重要手段，这种免疫响应广泛存在于从低等原核生物到高等哺乳动物当中（图1）。哺乳动物细胞利用多条天然免疫通路识别病原DNA并介导免疫应答；细菌则主要依赖“R-M系统”及“CRISPR-Cas系统”来去除入侵的噬菌体DNA。 图1：宿主细胞对病原DNA的免疫识别和应答 限制修饰（...
PCC7002, leveraging its endogenous type I-D CRISPR-Cas system. Utilizing this novel tool, we successfully deleted the glgA1 gene and iteratively edited the genome to obtain a double mutant of glgA1 and glgA2 genes. Additionally, large DNA fragments encompassing the entire type I-A (14 kb...
We used Pectobacterium atrosepticum, a Gram-negative phytopathogen, to study CRISPR-Cas regulation, since it contains a single Type I-F system. The CRP-cAMP complex activated the cas operon, increasing the expression of the adaptation genes cas1 and cas2-3 in addition to the genes encoding ...
The type I-F CRISPR–Cas system fromPseudomonas aeruginosa(PA14) provides sequence-specific elimination of invading DNA1,2. Recently, a paper published inCell Researchreported that this system targets mRNA through a non-canonical mechanism3. Here, we implement the proposed design rules for generating...