Repairing TALEN-mediated double-strand break by microhomology-mediated recombination in tobacco plastids generates abundant subgenomic DNA

Huang, Chih‐Hao; Liu, Yu-Chang; Shen, Jiayi; Lu, Fu I.; Shaw, Shyh-Yu; Huang, Hsiang‐Po; Chang, Ching Chih

doi:10.1016/j.plantsci.2021.111028

Cited by 8 publications

(21 citation statements)

References 57 publications

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“…Moreover, TALENs have been used to achieve multiplex genome-editing (MGE), for example, using a single TALEN pair to edit three homoeoalleles encoding mildew resistance locus proteins in hexaploid bread wheat [ 270 , 271 ]. In addition to nuclear genome editing mediated by TALENs, this technology has been successfully used to target and edit mitochondrial and chloroplast sequences (mitoTALENs and cpTALEN, respectively) [ 272 , 273 , 274 , 275 , 276 ] ( Figure 2 f,g).…”

Section: Molecular Toolsmentioning

confidence: 99%

Biotechnological Advances to Improve Abiotic Stress Tolerance in Crops

Villalobos-López

Arroyo-Becerra

Quintero-Jiménez

et al. 2022

IJMS

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The major challenges that agriculture is facing in the twenty-first century are increasing droughts, water scarcity, flooding, poorer soils, and extreme temperatures due to climate change. However, most crops are not tolerant to extreme climatic environments. The aim in the near future, in a world with hunger and an increasing population, is to breed and/or engineer crops to tolerate abiotic stress with a higher yield. Some crop varieties display a certain degree of tolerance, which has been exploited by plant breeders to develop varieties that thrive under stress conditions. Moreover, a long list of genes involved in abiotic stress tolerance have been identified and characterized by molecular techniques and overexpressed individually in plant transformation experiments. Nevertheless, stress tolerance phenotypes are polygenetic traits, which current genomic tools are dissecting to exploit their use by accelerating genetic introgression using molecular markers or site-directed mutagenesis such as CRISPR-Cas9. In this review, we describe plant mechanisms to sense and tolerate adverse climate conditions and examine and discuss classic and new molecular tools to select and improve abiotic stress tolerance in major crops.

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Section: Molecular Toolsmentioning

confidence: 99%

Biotechnological Advances to Improve Abiotic Stress Tolerance in Crops

Villalobos-López

Arroyo-Becerra

Quintero-Jiménez

et al. 2022

IJMS

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“…Upon creation of DSBs by the endonuclease fused to the TALE repeats (Figure 4), the cellular repair machinery often creates small insertions or deletions by NHEJ [178]. TALENs have also been successfully used to knock out genes in plastids and mitochondria [179][180][181]. In plant organellar genomes, however, no NHEJ by re-ligation of broken DNA double strands takes place and DNA repair appears to occur predominantly by HR [182].…”

Section: Genome Editing With Ddcbes In Plant Organellar Genomesmentioning

confidence: 99%

DNA base editing in nuclear and organellar genomes

et al. 2022

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“…Transcription activator-like effector nucleases (TALENs) play an important role in genome editing because of the high specificity, the feasibility of organellar targeting and their high efficiency in heterochromatin regions [ 20 , 21 , 22 , 23 , 24 , 25 ]. For instance, the use of TALENs fused with mitochondria localization signals in the N-terminus (mitoTALENs) could successfully restore male fertility in rice and rapeseed CMS varieties by knocking out cytoplasmic male sterility (CMS)-associated genes [ 21 ].…”

Section: Introductionmentioning

confidence: 99%

“…Bacterial cytidine deaminase fused to the DNA binding domains of TALENs and chloroplast transit peptide could homoplasmically substitute the targeted Cs to Ts in the plastid DNA of Arabidopsis, and the targeted base editing mutations could be inherited by their offspring [ 24 ]. Recently, we used a leaf-specific rbcS promoter to drive the expression of TALENs fused with a transit peptide to site-specifically caused double-strand DNA breaks in the rpoB gene of tobacco [ 25 ]. Both photosynthetically competent and incompetent transgenic tobacco lines were generated [ 25 ].…”

Section: Introductionmentioning

confidence: 99%

“…Recently, we used a leaf-specific rbcS promoter to drive the expression of TALENs fused with a transit peptide to site-specifically caused double-strand DNA breaks in the rpoB gene of tobacco [ 25 ]. Both photosynthetically competent and incompetent transgenic tobacco lines were generated [ 25 ]. In an albino line, the repair of damaged cpDNA through microhomology-mediated recombination caused the functional loss of the rpoB operon and resulted in the heterotrophic growth of the plant [ 25 ].…”

Section: Introductionmentioning

confidence: 99%

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A Transcriptomic Analysis of Tobacco Leaf with the Functional Loss of the Plastid rpoB Operon Caused by TALEN-Mediated Double-Strand Breakage

Liu

Huang

Chang

2022

Plants

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At least two sets of RNA polymerase (RNAP), nucleus (NEP)- and plastid (PEP)-encoded polymerases, recognizing distinct promoters exist in the plastids of land plants. Most plastid genes are regulated by multiple promoters with different strengths in their response to developmental stages and environmental cues. Recently, we applied chloroplast-targeted transcription activator-like effector nuclease (cpTALEN) technology to site-specifically cause double-strand DNA breaks in the rpoB gene of tobacco, which encodes the β-subunit of PEP. The repair of damaged chloroplast DNA (cpDNA) through microhomology-mediated recombination caused the functional loss of the rpoB operon and resulted in the heterotrophic growth of an albino plant. We conducted a genome-wide analysis of the steady state of gene expression in the leaf tissue of PEP-deficient tobacco by RNA-Seq and compared it with that of wild-type plants. The expression of NEP genes was up-regulated in PEP-deficient tobacco; in particular, the level of RpoT3 transcripts encoding the specifically plastid-targeted NEP was significantly increased. Alongside most housekeeping genes, NEP also plays an important role in the regulation of gene expression involved in photosynthesis. In contrast, alongside the photosynthesis-related genes, PEP also plays an important role in the regulation of gene expression involved in some housekeeping functions. Furthermore, the mitochondrial DNA copy number and the level of most mitochondrial protein-coding transcripts were slightly increased in PEP-deficient tobacco. The disruption of PEP function not only affected plastid gene expression, but also nuclear and mitochondrial gene expression. This study demonstrated the intercompartmental retrograde signaling in the regulation of gene expression.

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Repairing TALEN-mediated double-strand break by microhomology-mediated recombination in tobacco plastids generates abundant subgenomic DNA

Cited by 8 publications

References 57 publications

Biotechnological Advances to Improve Abiotic Stress Tolerance in Crops

Biotechnological Advances to Improve Abiotic Stress Tolerance in Crops

DNA base editing in nuclear and organellar genomes

A Transcriptomic Analysis of Tobacco Leaf with the Functional Loss of the Plastid rpoB Operon Caused by TALEN-Mediated Double-Strand Breakage

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