Large-scale discovery of induced point mutations with high-throughput TILLING.

By B. Till, S. Reynolds, E. Greene, Christine A. Codomo, L. Enns, Jessica E Johnson, C. Burtner, Anthony R. Odden, Kim Young, Nicholas E. Taylor and 3 more. Genome Research, 2003

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TILLING (Targeting Induced Local Lesions in Genomes) is a general reverse-genetic strategy that provides an allelic series of induced point mutations in genes of interest. High-throughput TILLING allows the rapid and low-cost discovery of induced point mutations in populations of chemically mutagenized individuals. As chemical mutagenesis is widely applicable and mutation detection for TILLING is dependent only on sufficient yield of PCR products, TILLING can be applied to most organisms. We have developed TILLING as a service to the Arabidopsis community known as the Arabidopsis TILLING Project (ATP). Our goal is to rapidly deliver allelic series of ethylmethanesulfonate-induced mutations in target 1-kb loci requested by the international research community. In the first year of public operation, ATP has discovered, sequenced, and delivered >1000 mutations in >100 genes ordered by Arabidopsis researchers. The tools and methodologies described here can be adapted to create similar facilities for other organisms.

Subfields: Molecular Biology, Plant Science

View Paper: https://www.semanticscholar.org/paper/77f729c3d2b9bce67ee3e5fcfd2231a6fa0f9e17

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Citations in the corpus

Genome Editing of Plants (2017, methodology): “Till et al. (2003) refined the TILLING process that enabled high throughput screening of Arabidopsis mutants by incorporation of a digestion step using the mismatch cleavage enzyme CEL I and visualization of any resulting DNA fragments via denaturing polyacrylamide gel electrophoresis.”
Comprehending crystalline β-carotene accumulation by comparing engineered cell models and the natural carotenoid-rich system of citrus (2011, neutral): “In order to find a SLAC1-deficient mutant of rice, a screening procedure called TILLING (Targeting Induced Local Lesions IN Genomes), whereby pinpoint mutations in individual genes can be detected using a PCR-based assay, was used (Till et al., 2003).”
Prospects for crop production under drought: research priorities and future directions (2005, methodology): “One powerful method is TILLING (targeting-induced local lesions in genomes) that allows the generation of targeted mutations, allowing gene candidates discovered by genomics-based experimentation to be used to directly alter trait components (Henikoff & Comai, 2003; Till et al., 2003a,b).”
The Arabidopsis Information Resource: Making and Mining the ‘Gold Standard’ Annotated Reference Plant Genome (2015, background): “…of thousands of insertion lines, knockdowns, point mutations, over-expressors and more have been donated to the stock centers and many of the insertion sites have been mapped to the genome (Alonso et al., 2003; Rosso et al ., 2003; Sessions et al ., 2002; Till et al ., 2003; Woody et al ., 2007).”
Spectrum of chemically induced mutations from a large-scale reverse-genetic screen in Arabidopsis. (2003, methodology): “…Our data set derives from accumulation of mutation procedures are described elsewhere (Till et al. 2003a,b). data generated by the Arabidopsis TILLING ProjectBriefly, seeds were mutagenized by soaking 10–20 hr in 20–40 over its first 18 months of operation (Till et al. 2003b).mm EMS and sown.”
Ethyl Methane Sulphonate (EMS) Induced Mutagenesis in Malaysian Rice (cv. MR219) for Lethal Dose Determination (2012, background): “In Arabidopsis, point mutation density can be as high as four mutations per Mb [3-5].”
Induced Mutations Unleash the Potentials of Plant Genetic Resources for Food and Agriculture (2013, background): “These include Arabidopsis [67–70]; rice [71–74]; maize [75]; wheat [76,77]; sugar beet [78]; barley [79]; soybean [80]; pea [81]; beans [82,83]; tomato [84] and in the vegetatively propagated banana [85].”