Patent
Multilayer genetic safety kill circuits based on single cas9 protein and multiple engineered grna in mammalian cells
TLDR
In this article, the authors describe a regulatory system composed of a CRISPR-associated nuclease and at least two distinct guide RNAs (gRNAs), which modulates cleavage and transcription, including repression and activation, in a mammalian cell such as a human cell.Abstract:
Aspects of the disclosure relate to synthetic regulatory systems composed of a multifunctional Cas [clustered regularly interspaced short palindromic repeat (CRISPR)-associated (Cas)] nuclease and at least two distinct guide RNAs (gRNAs). The synthetic regulatory system modulates cleavage and transcription, including repression and activation, in a mammalian cell such as a human cell.read more
Citations
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Patent
Adenosine nucleobase editors and uses thereof
David R. Liu,Nicole M. Gaudelli +1 more
TL;DR: In this article, the authors provide fusion proteins comprising a Cas9 (e.g., a Cas 9 nickase) domain and adenosine deaminases that deaminate adenosines in DNA, and in some embodiments, the fusion proteins further comprise a nuclear localization sequence (NLS), and/or an inhibitor of base repair, such as, a nuclease dead inosine specific nucolate (dISN).
Patent
Cas9 proteins including ligand-dependent inteins
David R. Liu,Kevin Davis +1 more
TL;DR: In this article, the authors provide compositions, methods, systems, and kits for controlling the activity of RNA-programmable endonucleases, such as Cas9, or for controlling proteins comprising a Cas9 variant fused to a functional effector domain.
Patent
Delivery of negatively charged proteins using cationic lipids
TL;DR: In this article, the authors present compositions, methods, strategies, kits, and systems for the delivery of negatively charged proteins, protein complexes, and fusion proteins using cationic polymers or lipids.
Patent
Editing of CCR5 receptor gene to protect against HIV infection
Juan Pablo Maianti,David R. Liu +1 more
TL;DR: In this paper, the authors present systems, compositions, and methods of introducing protective and/or loss-of-function variants of CCR5 and CCR2 for HIV infection and progression.
References
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Journal ArticleDOI
A programmable dual-RNA-guided DNA endonuclease in adaptive bacterial immunity.
Martin Jinek,Krzysztof Chylinski,Krzysztof Chylinski,Ines Fonfara,Michael H. Hauer,Jennifer A. Doudna,Emmanuelle Charpentier +6 more
TL;DR: This study reveals a family of endonucleases that use dual-RNAs for site-specific DNA cleavage and highlights the potential to exploit the system for RNA-programmable genome editing.
Journal ArticleDOI
Highly efficient Cas9-mediated transcriptional programming
Alejandro Chavez,Jonathan Scheiman,Suhani Vora,Benjamin W. Pruitt,Marcelle Tuttle,Eswar Prasad Ramachandran Iyer,Shuailiang Lin,Samira Kiani,Christopher D. Guzman,Daniel J. Wiegand,Dmitry Ter-Ovanesyan,Jonathan L. Braff,Noah Davidsohn,Benjamin E. Housden,Norbert Perrimon,Ron Weiss,John Aach,James J. Collins,George M. Church +18 more
TL;DR: An improved transcriptional regulator obtained through the rational design of a tripartite activator, VP64-p65-Rta (VPR), fused to nuclease-null Cas9 is described and demonstrated in activating endogenous coding and noncoding genes and stimulating neuronal differentiation of human induced pluripotent stem cells (iPSCs).
PatentDOI
Orthogonal cas9 proteins for rna-guided gene regulation and editing
TL;DR: In this article, the use of multiple orthogonal Cas9 proteins to simultaneously regulate corresponding genes or simultaneously and independently edit corresponding genes is described. But the authors do not consider the effect of the Cas9 protein on the expression of the target nucleic acid.
Journal ArticleDOI
Multiplex genome engineering in human cells using all-in-one CRISPR/Cas9 vector system
TL;DR: This work established a system for constructing all-in-one expression vectors containing multiple guide RNA expression cassettes and a Cas9 nuclease/nickase expression cassette that provides an efficient targeting strategy for multiplex genome/epigenome editing, simultaneous activation/repression of multiple genes, and beyond.
Journal ArticleDOI
Cas9 gRNA engineering for genome editing, activation and repression
Samira Kiani,Alejandro Chavez,Alejandro Chavez,Marcelle Tuttle,Richard N. Hall,Raj Chari,Dmitry Ter-Ovanesyan,Dmitry Ter-Ovanesyan,Jason Qian,Jason Qian,Benjamin W. Pruitt,Jacob Beal,Suhani Vora,Suhani Vora,Suhani Vora,Joanna Buchthal,Emma J. K. Kowal,Mohammad R. Ebrahimkhani,James J. Collins,Ron Weiss,George M. Church,George M. Church +21 more
TL;DR: In this paper, the length of Cas9-associated guide RNA (gRNA) was altered to control Cas9 nuclease activity and simultaneously perform genome editing and transcriptional regulation with a single Cas9 protein.
Related Papers (5)
High-throughput screens in mammalian cells using the CRISPR-Cas9 system.
Generation of a conditional analog-sensitive kinase in human cells using CRISPR/Cas9-mediated genome engineering
Tyler C. Moyer,Andrew J. Holland +1 more