CRISPR gene editing reaches inside plant cells' solar power factories
Mirzaee M, Best C, Wachowski EV, Maliga P
Crispr
Chloroplasts control photosynthesis and are passed down only through seeds, not pollen, so editing them precisely could one day mean hardier crops whose engineered traits can't drift into wild relatives.
Every green plant cell has tiny structures called chloroplasts that capture sunlight and turn it into food, and they carry their own small set of DNA separate from the main plant genome. Researchers figured out how to get the CRISPR gene-editing system, usually used on a plant's main DNA, to work inside these chloroplasts too, using tobacco plants as their test subject. This is an early but important step toward one day editing chloroplast DNA in food crops to improve things like stress tolerance or yield.
Key Findings
Researchers engineered tobacco chloroplasts to produce properly sized guide RNAs using an rRNA promoter and either a tRNA (trnG) or HDV ribozyme to define the RNA ends.
Cas9, delivered from the nucleus via a transit peptide, successfully entered chloroplasts and cut plastid DNA at the ndhA and rpoC1 genes, with breaks repaired via microhomology-mediated end joining (MMEJ), causing deletions.
Guide RNAs expressed from the plant's nucleus could be delivered into chloroplasts by fusing them to viroid RNA, suggesting a path to chloroplast editing without direct chloroplast transformation.
chevron_right Technical Summary
Scientists successfully used CRISPR gene-editing tools to make precise cuts in the DNA of chloroplasts, the tiny solar-powered factories inside plant cells, opening a new path to engineer traits like disease resistance or improved photosynthesis directly in these organelles.
Abstract Preview
Original paper
RNA-Guided Engineering of the Chloroplast Genome Enabled by Plastid-Expressed Guide RNAs.
Our goal is to develop RNA-guided engineering of the chloroplast genome using the CRISPR/Cas9 system. We designed chloroplast minigenes to obtain properly sized single guide RNAs (sgRNAs) in tobacc...
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Tobacco is the common name of several plants in the genus Nicotiana of the family Solanaceae, and the general term for any product prepared from the cured leaves of these plants. Seventy-nine species of tobacco are known, but the chief commercial crop is N. tabacum. The more potent variant N. rus...