公开(公告)号：US20140349400A1

公开(公告)日：2014-11-27

申请号：US14217426

申请日：2014-03-17

申请人： Noah Jakimo ,  Peter A. Carr ,  Joseph M. Jacobson

发明人： Noah Jakimo ,  Peter A. Carr ,  Joseph M. Jacobson

IPC分类号： C12N15/63

CPC分类号： C12N15/635 ,  C12N15/102

摘要： A self-reconfiguring genome uses a cassette having operons or DNA sequences that code for guide RNA, reverse transcriptase, donor RNA, and a CRISPR cleavage enzyme. A self-reconfiguring genome may be based on lambda recombineering of in situ generated oligonucleotides. A method for programmable self-modification of a cellular genome includes transcribing guide RNA from a self-reconfiguring cassette, associating the transcribed guideRNA with the CRISPR enzyme, intercalcating a region of complimentary sequence within an integration site of the genome, cutting upstream of a PAM site within the integration site; transcribing the donorRNA, translating donorRNA to double-stranded DNA, and recombining the double-stranded DNA via homologous recombination at the cut site of the integration site. A set of cascadable and multiplexable genetic logic gates with a universal RNA input/output based on single-strand annealing or non-homologous end joining, comprises transcription promoters or terminators, homologous regions, DNA sequences, RNA, and enzymes from the CRISPR system.

摘要翻译： 自重构基因组使用具有编码引导RNA，逆转录酶，供体RNA和CRISPR切割酶的操纵子或DNA序列的盒。 自重构基因组可以基于原位产生的寡核苷酸的λ重组。 用于细胞基因组的可编程自修饰的方法包括从自重构盒转录引导RNA，将转录的引导RNA与CRISPR酶缔合，在基因组的整合位点嵌入互补序列的区域，切割PAM的上游 整合网站内的网站; 转录供体RNA，将供体RNA翻译成双链DNA，并通过同源重组在整合位点的切割位点重组双链DNA。 一组具有基于单链退火或非同源末端连接的通用RNA输入/输出的可级联和可复用的遗传逻辑门包括来自CRISPR系统的转录启动子或终止子，同源区，DNA序列，RNA和酶。