Screening for CRISPR and TALENs Edits, Better with RibonucleoproteinsNovember 28, 2018
Researchers develop ribonucleoprotein (RNP)-based screening for genome modifications by CRISPR and TALENs because of limitations of existing methodologies for screening. These methods include polymerase chain reaction (PCR) with restriction enzyme digestion, T7EI cleavage assay, and sequencing, which are limited by sequence requirement, non-applicability in high ploidy species, non-detection of heterozygous mutations, and high cost.
PCR/RNP works by cleaving DNA sequences that are identical to the RNA in the RNP complex. The expected result of using this method is multiple bands for non-edited plants and intact single bands for edited plants for diploid or polyploid plants. Researcher Zhen Liang from University of Chinese Academy of Sciences and colleagues tested this method in hexaploid wheat and diploid rice that were edited using CRISPR and TALENs. Results showed higher sensitivity of the method than DNA sequencing, high applicability in hexaploid wheat, and ability to detect edited genes using TALENs. This method proved to be useful because of its low cost and short time span requirement.
For more information, read the article in Plant Biotechnology Journal.
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