Multiplexed genome engineering by Cas12a and CRISPR arrays encoded on single transcripts.
Nat Methods
; 16(9): 887-893, 2019 09.
Article
em En
| MEDLINE
| ID: mdl-31406383
ABSTRACT
The ability to modify multiple genetic elements simultaneously would help to elucidate and control the gene interactions and networks underlying complex cellular functions. However, current genome engineering technologies are limited in both the number and the type of perturbations that can be performed simultaneously. Here, we demonstrate that both Cas12a and a clustered regularly interspaced short palindromic repeat (CRISPR) array can be encoded in a single transcript by adding a stabilizer tertiary RNA structure. By leveraging this system, we illustrate constitutive, conditional, inducible, orthogonal and multiplexed genome engineering of endogenous targets using up to 25 individual CRISPR RNAs delivered on a single plasmid. Our method provides a powerful platform to investigate and orchestrate the sophisticated genetic programs underlying complex cell behaviors.
Texto completo:
1
Coleções:
01-internacional
Base de dados:
MEDLINE
Assunto principal:
Engenharia Genética
/
Genoma Humano
/
RNA Guia de Cinetoplastídeos
/
Endonucleases
/
Redes Reguladoras de Genes
/
Sistemas CRISPR-Cas
/
Edição de Genes
Limite:
Humans
Idioma:
En
Revista:
Nat Methods
Ano de publicação:
2019
Tipo de documento:
Article