The CRISPR/Cas9 system sheds new lights on the biology of protozoan parasites

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Abstract

The CRISPR/Cas9 system, a natural defence system of bacterial organisms, has recently been used to modify genomes of the most important protozoa parasites. Successful genome manipulations with the CRISPR/Cas9 system are changing the present view of genetics in parasitology. The application of this system offers a major chance to overcome the current restriction in culturing, maintaining and analysing protozoan parasites, and allows dynamic analysis of parasite genes functions, leading to a better understanding of pathogenesis. CRISPR/Cas9 system will have a significant influence on the process of developing novel drugs and treatment strategies against protozoa parasites.

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Most cited references 49

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Functional repair of CFTR by CRISPR/Cas9 in intestinal stem cell organoids of cystic fibrosis patients.

Gerald Schwank, Bon-Kyoung Koo, Valentina Sasselli … (2013)

Single murine and human intestinal stem cells can be expanded in culture over long time periods as genetically and phenotypically stable epithelial organoids. Increased cAMP levels induce rapid swelling of such organoids by opening the cystic fibrosis transmembrane conductor receptor (CFTR). This response is lost in organoids derived from cystic fibrosis (CF) patients. Here we use the CRISPR/Cas9 genome editing system to correct the CFTR locus by homologous recombination in cultured intestinal stem cells of CF patients. The corrected allele is expressed and fully functional as measured in clonally expanded organoids. This study provides proof of concept for gene correction by homologous recombination in primary adult stem cells derived from patients with a single-gene hereditary defect. Copyright © 2013 Elsevier Inc. All rights reserved.

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Genome Engineering of Drosophila with the CRISPR RNA-Guided Cas9 Nuclease

Scott Gratz, Alexander M. Cummings, Jennifer Nguyen … (2013)

We have adapted a bacterial CRISPR RNA/Cas9 system to precisely engineer the Drosophila genome and report that Cas9-mediated genomic modifications are efficiently transmitted through the germline. This RNA-guided Cas9 system can be rapidly programmed to generate targeted alleles for probing gene function in Drosophila.

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CRISPR/Cas9 in Genome Editing and Beyond

Haifeng Wang, Marie La Russa, Lei S Qi (2016)

The Cas9 protein (CRISPR-associated protein 9), derived from type II CRISPR (clustered regularly interspaced short palindromic repeats) bacterial immune systems, is emerging as a powerful tool for engineering the genome in diverse organisms. As an RNA-guided DNA endonuclease, Cas9 can be easily programmed to target new sites by altering its guide RNA sequence, and its development as a tool has made sequence-specific gene editing several magnitudes easier. The nuclease-deactivated form of Cas9 further provides a versatile RNA-guided DNA-targeting platform for regulating and imaging the genome, as well as for rewriting the epigenetic status, all in a sequence-specific manner. With all of these advances, we have just begun to explore the possible applications of Cas9 in biomedical research and therapeutics. In this review, we describe the current models of Cas9 function and the structural and biochemical studies that support it. We focus on the applications of Cas9 for genome editing, regulation, and imaging, discuss other possible applications and some technical considerations, and highlight the many advantages that CRISPR/Cas9 technology offers.

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Author and article information

Contributors

Maciej Grzybek:

ORCID: http://orcid.org/0000-0002-8780-0304

+ 4858 3491941 , grzybek.genetics@gmail.com

Aleksandra Golonko: olau.95@gmail.com

Aleksandra Górska: aleksandra.olejarska@up.lublin.pl

Klaudiusz Szczepaniak: k.o.szczepaniak@gmail.com

Aneta Strachecka: aneta.strachecka@up.lublin.pl

Anna Lass: anna.ls1@gumed.edu.pl

Paweł Lisowski: pawel.lisowski@mdc-berlin.de

Journal

Journal ID (nlm-ta): Appl Microbiol Biotechnol

Journal ID (iso-abbrev): Appl. Microbiol. Biotechnol

Title: Applied Microbiology and Biotechnology

Publisher: Springer Berlin Heidelberg (Berlin/Heidelberg )

ISSN (Print): 0175-7598

ISSN (Electronic): 1432-0614

Publication date (Electronic): 6 April 2018

Publication date PMC-release: 6 April 2018

Publication date (Print): 2018

Volume: 102

Issue: 11

Pages: 4629-4640

Affiliations

[1 ]ISNI 0000 0001 0531 3426, GRID grid.11451.30, Department of Tropical Parasitology, , Medical University of Gdansk, ; Powstania Styczniowego 9b, 81-519 Gdynia, Poland

[2 ]ISNI 0000 0000 9787 2307, GRID grid.446127.2, Department of Biotechnology, , Bialystok University of Technology, ; Wiejska 45E, 15-351, Bialystok, Poland

[3 ]ISNI 0000 0000 8816 7059, GRID grid.411201.7, Faculty of Veterinary Medicine, , University of Life Sciences in Lublin, ; Akademicka 13, 20-950 Lublin, Poland

[4 ]ISNI 0000 0000 8816 7059, GRID grid.411201.7, Faculty of Biology, Animal Sciences and Bioeconomy, , University of Life Sciences in Lublin, ; Akademicka 13, 20-950 Lublin, Poland

[5 ]ISNI 0000 0001 1210 151X, GRID grid.460378.e, Department of Molecular Biology, , Institute of Genetics and Animal Breeding PAS, ; Postepu 36A, 05-552 Jastrzebiec, Poland

[6 ]ISNI 0000 0001 1942 5154, GRID grid.211011.2, iPS Cell-Based Disease Modeling Group, Max-Delbrück-Center for Molecular Medicine (MDC) in the Helmholtz Association, ; Robert-Rössle-Str. 10, 13092 Berlin, Germany

Author information

Maciej Grzybek http://orcid.org/0000-0002-8780-0304

Article

Publisher ID: 8927

DOI: 10.1007/s00253-018-8927-3

PMC ID: 5954013

PubMed ID: 29626235

SO-VID: 79bb2f8d-52de-4722-ab9f-dfe40bfe4bd7

License:

Open Access This article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made.

History

Date received : 2 March 2018

Date accepted : 6 March 2018

Funding

Funded by: FundRef http://dx.doi.org/10.13039/501100004281, Narodowe Centrum Nauki;

Award ID: 2012/07/N/NZ9/02060

Award ID: 2016/22/M/NZ2/00548

Award Recipient : Maciej Grzybek

Funded by: FundRef http://dx.doi.org/10.13039/501100004569, Ministerstwo Nauki i Szkolnictwa Wyższego;

Award ID: 428/STYP/11/2016

Award Recipient : Maciej Grzybek

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ScienceOpen disciplines: Biotechnology

Keywords: crispr,cas9,genetic modification,protozoa,apicomplexa,genome editing

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ScienceOpen disciplines: Biotechnology

Keywords: crispr, cas9, genetic modification, protozoa, apicomplexa, genome editing

The CRISPR/Cas9 system sheds new lights on the biology of protozoan parasites

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Abstract

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CRISPR/Cas9 editing in human blood

Most cited references 49

Functional repair of CFTR by CRISPR/Cas9 in intestinal stem cell organoids of cystic fibrosis patients.

Genome Engineering of Drosophila with the CRISPR RNA-Guided Cas9 Nuclease

CRISPR/Cas9 in Genome Editing and Beyond

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