公开(公告)号：WO2016183438A1

公开(公告)日：2016-11-17

申请号：PCT/US2016/032348

申请日：2016-05-13

Applicant: MASSACHUSETTS INSTITUTE OF TECHNOLOGY

Inventor： LU, Timothy, Kuan-ta ,  PERLI, Samuel, David ,  CUI, Hao

IPC: C12N15/00 ,  C12N15/63 ,  G06N3/12

CPC classification number: C12N15/63 ,  C12N15/102 ,  C12N15/111 ,  C12N2310/20 ,  G06N3/12

Abstract: The present disclosure is directed, in some embodiments, to engineered nucleic acids comprising a promoter operably linked to a nucleotide sequence encoding a guide ribonucleic acid (gRNA) that comprises a specificity determining sequence (SDS) and a protospacer adjacent motif (PAM). The present disclosure is directed, in some embodiments, to cells comprising, vectors comprising, and methods of producing the engineered nucleic acids.

Abstract translation: 在一些实施方案中，本公开涉及包含与编码引导核糖核酸（gRNA）的核苷酸序列可操作地连接的启动子的工程化核酸，其包含特异性测定序列（SDS）和原原理邻近基序（PAM）。 在一些实施方案中，本公开涉及包含载体的细胞，以及生产工程化核酸的方法。

公开(公告)号：WO2015153940A1

公开(公告)日：2015-10-08

申请号：PCT/US2015/024196

申请日：2015-04-03

Applicant: MASSACHUSETTS INSTITUTE OF TECHNOLOGY

Inventor： LU, Timothy, Kuan-Ta ,  NISSIM, Lior ,  PERLI, Samuel, David

IPC: C12N15/63

CPC classification number: C12N15/113 ,  C12N9/22 ,  C12N15/111 ,  C12N15/63 ,  C12N15/85 ,  C12N2310/128 ,  C12N2310/141 ,  C12N2310/20 ,  C12N2310/51 ,  C12N2330/51 ,  C12N2830/42 ,  C12N2830/60 ,  C12N2830/85 ,  C12Y301/00

Abstract: Various aspects and embodiments of the present disclosure relate to methods and compositions that combine multiple mammalian RNA regulatory strategies, including RNA triple helix structures, introns, microRNAs, and ribozymes with Cas-based CRISPR trascription factors and ribonuclease -based RNA processing in human cells. The methods and compositions of the present disclosure, in some embodiments, enable multiplexed production of proteins and multiple guide RNAs from a single compact RNA-polymerase-II-expressed transcript for efficient modulation of synthetic constructs and endogenous human promoters.

Abstract translation: 本公开的各个方面和实施方案涉及将多种哺乳动物RNA调控策略（包括RNA三螺旋结构，内含子，微RNA和核酶）与基于Cas-based CRISPR trascription因子和基于核糖核酸酶的RNA加工在人类细胞中的方法和组合物。 在一些实施方案中，本公开的方法和组合物使得能够从单个紧凑的RNA聚合酶-II表达的转录物多肽产生蛋白质和多个引导RNA，以有效调节合成构建体和内源性人启动子。