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Title: Orthogonal Cas9 proteins for RNA-guided gene regulation and editing

Abstract

Methods of modulating expression of a target nucleic acid in a cell are provided including use of multiple orthogonal Cas9 proteins to simultaneously and independently regulate corresponding genes or simultaneously and independently edit corresponding genes.

Inventors:
; ;
Publication Date:
Research Org.:
President and Fellows of Harvard College, Cambridge, MA (United States)
Sponsoring Org.:
USDOE
OSTI Identifier:
1346066
Patent Number(s):
9,587,252
Application Number:
14/674,895
Assignee:
President and Fellows of Harvard College CHO
DOE Contract Number:
FG02-02ER63445
Resource Type:
Patent
Resource Relation:
Patent File Date: 2015 Mar 31
Country of Publication:
United States
Language:
English
Subject:
59 BASIC BIOLOGICAL SCIENCES

Citation Formats

Church, George M., Esvelt, Kevin, and Mali, Prashant. Orthogonal Cas9 proteins for RNA-guided gene regulation and editing. United States: N. p., 2017. Web.
Church, George M., Esvelt, Kevin, & Mali, Prashant. Orthogonal Cas9 proteins for RNA-guided gene regulation and editing. United States.
Church, George M., Esvelt, Kevin, and Mali, Prashant. Tue . "Orthogonal Cas9 proteins for RNA-guided gene regulation and editing". United States. doi:. https://www.osti.gov/servlets/purl/1346066.
@article{osti_1346066,
title = {Orthogonal Cas9 proteins for RNA-guided gene regulation and editing},
author = {Church, George M. and Esvelt, Kevin and Mali, Prashant},
abstractNote = {Methods of modulating expression of a target nucleic acid in a cell are provided including use of multiple orthogonal Cas9 proteins to simultaneously and independently regulate corresponding genes or simultaneously and independently edit corresponding genes.},
doi = {},
journal = {},
number = ,
volume = ,
place = {United States},
year = {Tue Mar 07 00:00:00 EST 2017},
month = {Tue Mar 07 00:00:00 EST 2017}
}

Patent:

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  • Methods of modulating expression of a target nucleic acid in a cell are provided including introducing into the cell a first foreign nucleic acid encoding one or more RNAs complementary to DNA, wherein the DNA includes the target nucleic acid, introducing into the cell a second foreign nucleic acid encoding a nuclease-null Cas9 protein that binds to the DNA and is guided by the one or more RNAs, introducing into the cell a third foreign nucleic acid encoding a transcriptional regulator protein or domain, wherein the one or more RNAs, the nuclease-null Cas9 protein, and the transcriptional regulator protein ormore » domain are expressed, wherein the one or more RNAs, the nuclease-null Cas9 protein and the transcriptional regulator protein or domain co-localize to the DNA and wherein the transcriptional regulator protein or domain regulates expression of the target nucleic acid.« less
  • Cited by 13
  • Cited by 13
  • SP6 bacteriophage RNA polymerase is produced by cultivating a new microorganism (particularly new strains of Escherichia coli) harboring a plasmid that carries SP6 bacteriophage RNA polymerase gene and recovering SP6 bacteriophage RNA polymerase from the culture broth. SP6 bacteriophage RNA polymerase gene is provided as are new microorganisms harboring a plasmid that carries SP6 bacteriophage RNA polymerase gene.
  • The commonly cited figure of 10{sup 5} genes in the human genome represents a tremendous underestimate of our capacity to generate distinct gene products with unique functions. Our cells possess an impressive collection of tools for altering the products of a single gene to create a variety of proteins. The different gene products may have related but distinct functions, allowing cells of different types or at different developmental stages to fine-tune their patterns of gene expression. These tools may act in the cytoplasm, as when proteins undergo post-translational modifications, or in the nucleus, in the processing of pre-mRNA. Two formsmore » of intranuclear fine-tuning are well established and widely studied: alternative splicing of pre-mRNAs and alternative polyadenylation site selection. In recent years it has become clear that cells possess yet another tool to create RNA sequence diversity, mRNA editing. The term {open_quotes}editing{close_quotes} is applied to posttranscriptional modifications of a purine or pyrimidine, which alter an mRNA sequence as it is read, for example, by ribosomes. Covalent changes to the structure of nucleotide bases are well known to occur on tRNA and rRNA molecules, but such changes in mRNA sequence are novel in that they have the capacity to change specific protein sequences. 43 refs., 1 fig.« less