Prashant Yadav

CRISPR/Cas9: a promising way to exploit genetic variation in plants

Creation of variation in existing gene pool of crop plants is the foremost requirement in crop improvement programmes. Genome editing is a tool to produce knock out of target genes either by introduction of insertion or by deletion that disrupts the function of a specific gene. The CRISPR/Cas9 (clustered regularly interspaced short palindromic repeats/CRISPR-associated protein 9) system is the most recent addition to the toolbox of sequence-specific nucleases that includes ZFNs and TALENs. The CRISPR/Cas9 system allows targeted cleavage of genomic DNA guided by a small noncoding RNA, resulting in gene modifications by both non-homologous end joining and homology-directed repair mechanisms. Here, we present an overview of mechanisms of CRISPR, its potential roles in creating variation in germplasm and applications of this novel interference pathway in crop improvement. The availability of the CRISPR/Cas9 system holds promise in facilitating both forward and reverse genetics and will enhance research in crops that lack genetic resources.

Recent Perspective of Next Generation Sequencing: Applications in Molecular Plant Biology and Crop Improvement

Nearly after three decades of Sanger sequencing method dominating the field of nucleotide sequencing, the new age of novel sequencing techniques commenced with 454-pyrosequencing in the year 2005 and thus started the era of next generation sequencing techniques. Since then many other novel sequencing techniques with increased accuracy, simplicity and cost-effectiveness have come up and called next to next generation sequencing techniques or third generation sequencing techniques. The scientific knowledge generated from next generation sequencing techniques has transformed the field of structural and functional genomic studies in different crop plants. In this review, various next generation sequencing techniques are described, with their applications and future prospective.