Genetic tools and techniques for Chlamydomonas reinhardtii

Abstract

The development of tools has always been a major driving force for the advancement of science. Optical microscopes were the first instruments that allowed discovery and descriptive studies of the subcellular features of microorganisms. Although optical and electron microscopes remained at the forefront of microbiological research tools since their inventions, the advent of molecular genetics brought about questions which had to be addressed with new "genetic tools". The unicellular green microalgal genus Chlamydomonas, especially the most prominent species C. reinhardtii, has become a frequently used model organism for many diverse fields of research and molecular genetic analyses of C. reinhardtii, as well as the available genetic tools and techniques, have become increasingly sophisticated throughout the last decades. The aim of this review is to provide an overview of the molecular key features of C. reinhardtii and summarize the progress related to the development of tools and techniques for genetic engineering of this organism, from pioneering DNA transformation experiments to state-of-the-art techniques for targeted nuclear genome editing and high-throughput screening approaches.

SUMO Protease SMT7 Modulates Ribosomal Protein L30 and Regulates Cell-Size Checkpoint Function

“…The unicellular green alga Chlamydomonas reinhardtii (Chlamydomonas) has a well-characterized haploid genetic system (Harris, 1989;Harris and Editor, 2009), sequenced genome (Merchant et al, 2007), relative paucity of gene duplications, a well-equipped molecular toolkit (Leon-Banares et al, 2004;Jinkerson and Jonikas, 2015;Mussgnug, 2015;Shin et al, 2016;Greiner et al, 2017), and recently added mutant libraries (Zhang et al, 2014;Li et al, 2016;Breker et al, 2018) that make it a unique microbial model for cell-size control and cell-cycle studies in the plant superkingdom (Tulin and Cross, 2014;Cross and Umen, 2015;Umen, 2018).…”

Time-resolved transcriptome analysis and lipid pathway reconstruction of the oleaginous green microalga Monoraphidium neglectum reveal a model for triacylglycerol and lipid hyperaccumulation

“…In the context of biodiesel production however, microalgal lipid productivity needs to be improved for overall economic feasibility [ 3 – 5 ]. A valuable strategy to reach this goal is metabolic engineering [ 6 , 7 ]. However, the current understanding of the algal lipid metabolism is still incomplete, although it has been progressively investigated in the model green alga Chlamydomonas reinhardtii [ 8 ].…”

Effect of green and red light in lipid accumulation and transcriptional profile of genes implicated in lipid biosynthesis in Chlamydomonas reinhardtii

“…23 Chlamydomonas reinhardtii is one of the most important models for biofuel production, because some of these microalgae offer advantages such as (a) fast growth rate, (b) whole-genome sequence availability, (c) easy genetic manipulation, (d) generation of stable transgenic lines over short periods, (e) confined cultivation devices to reduce environmental pollution, (f) facultative autotrophic and heterotrophic organism, and (8) no pollen formation, hence there is no risk of introducing transgenes in food crops. 24,25 To our best knowledge, there are no reports of the effect of red and green light in lipid accumulation in C. reinhardtii. In this work, we evaluate the behavior of this microalga under these two wavelengths (550 and 650 nm).…”