Compositions and methods for the expression of crispr guide rnas using the h1 promoter

• Main Authors: ZACK DONALD, RANGANATHAN VINOD
• Format: Patent
• Language: Chinese; English
• Published: 13.06.2017
• Subjects: BEER; BIOCHEMISTRY; CHEMISTRY; COMPOSITIONS THEREOF; CULTURE MEDIA; ENZYMOLOGY; METALLURGY; MICROBIOLOGY; MICROORGANISMS OR ENZYMES; MUTATION OR GENETIC ENGINEERING; PROPAGATING, PRESERVING OR MAINTAINING MICROORGANISMS; SPIRITS; VINEGAR; WINE

The presently disclosed subject matter provides compositions and methods for the expression of CRISPR guide RNAs using the HI promoter. In particular, compositions and methods are provided for the use of the HI promoter to express CRISPR guide RNA (gRNA) with altered specificity of the 5' nucleotide, as well as use of the HI promoter sequence as a bidirectional promoter to express Cas9 nuclease and the gRNA simultaneously. Compositions and methods are also provided for the expression and regulation of gRNA expression in vivo through the use of RNA ribozymes and regulatable aptazymes. 本公开主题提供用于使用H1启动子表达CRISPR向导RNA的组合物和方法。具体而言，提供了组合物和方法，其用于使用H1启动子来表达具有5'核苷酸的改变特异性的CRISPR向导RNA(gRNA)，以及使用H1启动子序列作为双向启动子来同时表达Cas9核酸酶和gRNA。还提供了组合物和方法，其通过使用RNA核酶和可调节的适体酶在体内表达和调节gRNA表达。