CRISPR/Cas based genome editing in microalgae
- Authors: Virolainen P.A.1, Chekunova E.M.1
-
Affiliations:
- Saint Petersburg State University
- Issue: Vol 21 (2023): Спецвыпуск
- Pages: 30-31
- Section: Genetically modified organism. The Нistory, Achivements, Social and Environmental Riscs
- URL: https://journals.eco-vector.com/ecolgenet/article/view/568609
- DOI: https://doi.org/10.17816/ecogen568609
- ID: 568609
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Abstract
CRISPR/Cas systems are presently the most attractive genome editing technology, that is widely used for genetic engineering of various crops and industrial microorganisms. Currently, application of the CRISPR/Cas based genome editing promises advances in microalgae biotechnology aimed at boosting the output of biofuels and valuable bioactive compounds. However, algae remain relatively complex objects for genetic manipulation [1]. The main problems are associated with the need of a species-oriented approach when creating a transformation toolbox due to the peculiarities in the structure of membranes and the cell wall of a particular taxon. The proper selection and design of a CRISPR construct is also required due to the possible presence of a powerful silencing system against introduced genetic constructs in the cell. These difficulties explain the low efficiency of microalgae transformation and the meager list of successfully edited species [1, 2].
The first instance of genome editing in microalgae using CRISPR/Cas was reported in Chlamydomonas reinhardtii P.A. Dang [3]. To date, four transformation methods (Agrobacterium-mediated, particle bombardment, glass beads agitation, electroporation) have been successfully used for editing (knock-in and knock-out) the C. reinhardtii genome with two types of CRISPR constructs (plasmid and ribonucleoprotein). The developed protocols make it possible to achieve high efficiency of genomic editing — for example, in our study it varied from 10.6% to 68.8% [4]. These benefits along with completely sequenced genome, well-studied genetics, accessibility and haplontic life cycle makes C. reinhardtii an outstanding model organism for CRISPR/Cas application in microalgae research [5].
Full Text
CRISPR/Cas systems are presently the most attractive genome editing technology, that is widely used for genetic engineering of various crops and industrial microorganisms. Currently, application of the CRISPR/Cas based genome editing promises advances in microalgae biotechnology aimed at boosting the output of biofuels and valuable bioactive compounds. However, algae remain relatively complex objects for genetic manipulation [1]. The main problems are associated with the need of a species-oriented approach when creating a transformation toolbox due to the peculiarities in the structure of membranes and the cell wall of a particular taxon. The proper selection and design of a CRISPR construct is also required due to the possible presence of a powerful silencing system against introduced genetic constructs in the cell. These difficulties explain the low efficiency of microalgae transformation and the meager list of successfully edited species [1, 2].
The first instance of genome editing in microalgae using CRISPR/Cas was reported in Chlamydomonas reinhardtii P.A. Dang [3]. To date, four transformation methods (Agrobacterium-mediated, particle bombardment, glass beads agitation, electroporation) have been successfully used for editing (knock-in and knock-out) the C. reinhardtii genome with two types of CRISPR constructs (plasmid and ribonucleoprotein). The developed protocols make it possible to achieve high efficiency of genomic editing — for example, in our study it varied from 10.6% to 68.8% [4]. These benefits along with completely sequenced genome, well-studied genetics, accessibility and haplontic life cycle makes C. reinhardtii an outstanding model organism for CRISPR/Cas application in microalgae research [5].
About the authors
Pavel A. Virolainen
Saint Petersburg State University
Email: s.pasha98@yandex.ru
ORCID iD: 0000-0001-5918-9395
phd student
Russian Federation, Saint PetersburgElena M. Chekunova
Saint Petersburg State University
Author for correspondence.
Email: elena_chekunova@mail.ru
ORCID iD: 0000-0001-8942-4771
dr. sci. (biol.), senior teacher
Russian Federation, Saint PetersburgReferences
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