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.

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