Mar 27, 2018

Assessment of two CRISPR-Cas9 genome editing protocols for rapid generation of Trypanosoma cruzi gene knockout mutants

International Journal for Parasitology
Gabriela Assis Burle-CaldasSantuza M R Teixeira

Abstract

CRISPR/Cas9 technology has been used to edit genomes in a variety of organisms. Using the GP72 gene as a target sequence, we tested two distinct approaches to generate Trypanosoma cruzi knockout mutants using the Cas9 nuclease and in vitro transcribed single guide RNA. Highly efficient rates of disruption of GP72 were achieved either by transfecting parasites stably expressing Streptococcus pyogenes Cas9 with single guide RNA or by transfecting wild type parasites with recombinant Staphylococcus aureus Cas9 previously associated with single guide RNA. In both protocols, we used single-stranded oligonucleotides as a repair template for homologous recombination and insertion of stop codons in the target gene.

  • References
  • Citations5

References

  • We're still populating references for this paper, please check back later.
  • References
  • Citations5

Citations

Mentioned in this Paper

Transfection
Genome
Genes
CRISPR-Cas Systems
Gene Editing
Knock-out
Candidate Disease Gene
Codon, Terminator
DNA, Protozoan
Recombination, Genetic

Related Feeds

CRISPR (general)

Clustered regularly interspaced short palindromic repeats (CRISPR) are DNA sequences in the genome that are recognized and cleaved by CRISPR-associated proteins (Cas). CRISPR-Cas system enables the editing of genes to create or correct mutations. Discover the latest research on CRISPR here.

CRISPR Ribonucleases Deactivation

CRISPR-Cas system enables the editing of genes to create or correct mutations. This feed focuses on mechanisms that underlie deactivation of CRISPR ribonucleases. Here is the latest research.

CRISPR for Genome Editing

Genome editing technologies enable the editing of genes to create or correct mutations. Clustered regularly interspaced short palindromic repeats (CRISPR) are DNA sequences in the genome that are recognized and cleaved by CRISPR-associated proteins (Cas). Here is the latest research on the use of CRISPR-Cas system in gene editing.

Related Papers

Molecular and Biochemical Parasitology
Gabriela Assis Burle-CaldasSantuza M R Teixeira
Current Opinion in Biotechnology
Khaoula BelhajVladimir Nekrasov
Current Opinion in Biotechnology
Jonathan L Schmid-Burgk
© 2020 Meta ULC. All rights reserved