Sequential Activation of Guide RNAs to Enable Successive CRISPR-Cas9 Activities.
Mol Cell
; 81(2): 226-238.e5, 2021 01 21.
Article
in En
| MEDLINE
| ID: mdl-33378644
ABSTRACT
Currently, either highly multiplexed genetic manipulations can be delivered to mammalian cells all at once or extensive engineering of gene regulatory sequences can be used to conditionally activate a few manipulations. Here, we provide proof of principle for a new system enabling multiple genetic manipulations to be executed as a preprogrammed cascade of events. The system leverages the programmability of the S. pyogenes Cas9 and is based on flexible arrangements of individual modules of activity. The basic module consists of an inactive single-guide RNA (sgRNA)-like component that is converted to an active state through the effects of another sgRNA. Modules can be arranged to bring about an algorithmic program of sequential genetic manipulations without the need for engineering cell-type-specific promoters or gene regulatory sequences. With the expanding diversity of available tools that use spCas9, this sgRNA-based system provides multiple levels of interfacing with mammalian cell biology.
Key words
Full text:
1
Collection:
01-internacional
Database:
MEDLINE
Main subject:
RNA, Guide, Kinetoplastida
/
Clustered Regularly Interspaced Short Palindromic Repeats
/
CRISPR-Cas Systems
/
Gene Editing
/
CRISPR-Associated Protein 9
Limits:
Animals
/
Humans
Language:
En
Journal:
Mol Cell
Journal subject:
BIOLOGIA MOLECULAR
Year:
2021
Document type:
Article