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citations
Cited by 17 publications
(6 citation statements)
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References 39 publications
(6 reference statements)
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“…(2019) used gene‐specific primers and found that 26 alpha‐gliadins were expressed in the developing endosperm of CS. Note that the number of genes varies between wheat varieties, and that expression levels vary among genes and between varieties (Shewry and Lookhart, 2003; Salentijn et al ., 2009; Noma et al ., 2019; Jouanin et al ., 2020a).…”
Section: Introductionmentioning
confidence: 99%
“…Thus, an accurate and precise CN measurement is critical for lines selection, being single or low CN generally desired [38]. Many studies have been carried out to compare the performances of qPCR and ddPCR in view of improving detection and quantification methods intended for those laboratories committed in official GMO control [15,[39][40][41][42][43][44]. In general, they argued in favor to ddPCR, being this technique insensitive to PCR inhibiting components often present in complex matrices and not dependent on calibration with standard curves obtained from certified reference materials for quantification.…”
Section: Discussionmentioning
confidence: 99%
“…An approximation proposed by Jouanin et al uses droplet digital PCR (ddPCR) to identify small (1–50 bp) and large (>300 bp) InDels in wheat α-gliadin genes targeted with CRISPR/Cas [ 45 ]. However, the authors also stated the need to further perform deep sequencing to characterize these mutations.…”
Section: Discussionmentioning
confidence: 99%
“…Following a similar genome editing approach, using guide RNAs targeting conserved domains in a-gliadin and c-gliadin gene families simultaneously, mutant lines with a significant reduction in both agliadin and c-gliadins were obtained (Jouanin et al 2019). Because a-gliadin genes are located in tandem arrays, genome editing targeting conserved regions within these genes simultaneously may result in partial or complete deletion of neighboring gliadin genes copies, and evidence for that effect was also obtained (Jouanin et al 2020a). The low-gluten wheat lines obtained in these studies provide a basis for subsequent editing studies to further reduce the epitope load in these wheat lines and to ultimately generate hypoimmunogenic-gluten wheat varieties (Jouanin et al 2020b).…”
Section: Genome Editing and Polyploid Cropsmentioning
confidence: 99%
“…While dPCR applications in genome-edited plant cells are still limited, the methodology clearly provides novel opportunities to quantitate small genome edits or low copy number targets in bulk populations of edited cells. Recently, Jouanin et al (2020) used dPCR mutation assays to detect indels (1 to 50 bp) and large deletions ( >300 bp) in wheat and concluded that dPCR is suitable for high-throughput screening of copy number variation and gene editing-induced mutations in large gene families. Consonant with this are the findings of Peng et al (2020) who developed a duplexed dPCR-based method for the detection and evaluation of gene editing frequencies in rice and allotetraploid rapeseed.…”
Section: Digital Pcrmentioning
confidence: 99%