Discovery and characterization of novel type I-D CRISPR-guided transposons identified among diverse Tn7-like elements in cyanobacteria.
Discovery and characterization of novel type I-D CRISPR-guided transposons identified among diverse Tn7-like elements in cyanobacteria.
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新型I型I型CRISPR引导的转座子的发现和表征在蓝细菌中不同的TN7样元素中鉴定出来。
DOI：
10.1093/nar/gkac1216
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发表时间：
2023-01-25
影响因子：
14.9
通讯作者：
Peters, Joseph E.
中科院分区：
文献类型：
--
作者：
Hsieh, Shan-Chi;Peters, Joseph E.
CRISPR-Cas防御系统已经被Tn 7家族细菌转座子天然地用于指导RNA定向转座。我们发现蓝藻基因组富含Tn 7样元件，包括大多数已知的指导RNA定向转座子，V-K，I-B1和I-B2型CRISPR-Cas系统。我们发现并表征了I-D型CRISPR-Cas系统的一个实例，该系统天然地被选择用于指导RNA定向转座。发现了I-D亚型特有的多种新适应，包括Cas 10核酸酶的自然失活。I-D型CRISPR-Cas转座系统在向导RNA长度要求方面显示出灵活性，并且可以被工程化以与基于核酶的自加工向导RNA一起发挥功能，从而在异源系统中去除对Cas 6的要求。I-D型CRISPR-Cas转座子还具有天然融合的转座酶蛋白，其对于剪切-粘贴转座具有功能。I-D型系统的多个属性为未来的基因编辑工作提供了独特的可能性。我们的生物信息学分析还揭示了对Tn 7样元素进化的更广泛理解。在蓝藻中的Tn 7样元件和趋同进化的多个例子中，确定了广泛的靶向系统交换，包括靶向整合到自然转化所需的基因中的系统。在蓝藻中发现并表征了用于RNA引导转座的I-D型CRISPR-Cas系统的共选择。CRISPR-Cas共选择发生，Cas 10 d核酸酶活性丧失和间隔区获得。
CRISPR-Cas defense systems have been naturally coopted for guide RNA-directed transposition by Tn7 family bacterial transposons. We find cyanobacterial genomes are rich in Tn7-like elements, including most of the known guide RNA-directed transposons, the type V-K, I-B1, and I-B2 CRISPR-Cas based systems. We discovered and characterized an example of a type I-D CRISPR-Cas system which was naturally coopted for guide RNA-directed transposition. Multiple novel adaptations were found specific to the I-D subtype, including natural inactivation of the Cas10 nuclease. The type I-D CRISPR-Cas transposition system showed flexibility in guide RNA length requirements and could be engineered to function with ribozyme-based self-processing guide RNAs removing the requirement for Cas6 in the heterologous system. The type I-D CRISPR-Cas transposon also has naturally fused transposase proteins that are functional for cut-and-paste transposition. Multiple attributes of the type I-D system offer unique possibilities for future work in gene editing. Our bioinformatic analysis also revealed a broader understanding of the evolution of Tn7-like elements. Extensive swapping of targeting systems was identified among Tn7-like elements in cyanobacteria and multiple examples of convergent evolution, including systems targeting integration into genes required for natural transformation. Cooption of a type I-D CRISPR-Cas system for RNA-guided transposition was discovered in cyanobacteria and characterized. CRISPR-Cas cooption occurred with loss of Cas10d nuclease activity and spacer acquisition.
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影响因子：
14.9
作者：
Letunic I;Bork P
通讯作者：
Bork P
影响因子：
64.8
作者：
Halpin-Healy, Tyler S.;Klompe, Sanne E.;Fernandez, Israel S.
通讯作者：
Fernandez, Israel S.
影响因子：
2.8
作者：
Khlebnikov, A;Datsenko, KA;Keasling, JD
通讯作者：
Keasling, JD
影响因子：
64.5
作者：
BAINTON, R;GAMAS, P;CRAIG, NL
通讯作者：
CRAIG, NL
影响因子：
16
作者：
Klompe SE;Jaber N;Beh LY;Mohabir JT;Bernheim A;Sternberg SH
通讯作者：
Sternberg SH