Targeted Mutagenesis in Hexaploid Bread Wheat Using the TALEN and CRISPR/Cas Systems

Abstract

The use of sequence-specific transcription activator-like effector nucleases (TALENs) and the clustered regularly interspaced short palindromic repeats-associated system (CRISPR/Cas9) have provided powerful reverse genetic approaches to the targeted modification of genomes in numerous organisms. Both systems have been employed to generate loss-of-function alleles in bread wheat, by targeting multiple and single copies of genes. Here we present protocols for modifying the wheat genome using the two systems. The protocols include the design of TALEN and CRISPR/Cas9 target sites and their construction, evaluation of their activities in protoplasts, transformation of plants, and mutation screening.

Maize Lethal Necrosis disease: review of molecular and genetic resistance mechanisms, socio-economic impacts, and mitigation strategies in sub-Saharan Africa

“…Large effect QTLs (qMCMV6-155.6, qMCMV6-156.5 and qMCMV10-135.8) identified in KS23-5, KS23-6 and DR-derived populations are controlled by recessive genes [ 94 ] that may be amenable to gene editing. With the recent additions of new tools like base editing [ 167 ] and prime editing [ 168 , 169 ], multiple genes from both major and minor QTLs can be targeted to pyramid high-level of resistance to MLN [ 170 ].…”