Study on editing tools of Plasmodium berghei Cas9 genome

Malaria, caused by infection of Plasmodium parasites, remains a world-wide public health burden. Although the genomes of many malaria parasites have been sequenced, we still do not know the functions of approximately half of the genes in the genomes. Studying gene function has become the focus of many studies; however, editing genes in malaria parasite genomes is still inefficient. As an emerging method, clustered regularly interspersed short palindromic repeats (CRISPR) /Cas9 has been applied in genome editing at may model organisms. Two key components in this system, nuclease Cas9 and guide RNA sgRNA, form Cas9/sgRNA complex to perform the targeted DNA recognition and double strand break. Previous work has established the CRISPR/Cas9-meditaed genome editing method in a rodent malaria parasite Plasmodium yoelii. However, does this method perform well in another rodent malaria parasite model Plasmodium berghei is unknown? Recent work has established the expression and nuclear localization of Cas9 driven by 2A peptide with hDHFR selector marker in P. berghei. U6 snRNA promoter derived from P. yoelii is capable of driving transcription of sgRNA in P. berghei. It has been further demonstrated that Cas9/sgRNA is able to induce DNA double-strand breaks with specificity for targeted sites and that these breaks can be repaired efficiently by homologous recombination. By supplying engineered homologous repair templates, the project generated targeted deletion, tag addition, and nucleotide replacement targeting to endogenous gene in P. berghei.

Genetics and Genomics

Jan 2016 — Dec 2018

$29,971

China