ABSTRACT
The enigma surrounding life always makes humans inquisitive towards understanding their mechanism. With the developments in the past decades, science has witnessed rapid strides in terms of high-throughput technologies, including sequencing, genotyping, targeted genome technologies and computational methods. This played a major role towards probing various molecular mechanisms in plants. The availability of the draft genome sequences of the plant genomes enhances the scope for systematic studies for understanding the functions of the genes. Identifying the function of a particular gene is crucial to understand the underlying mechanism in the life process. Thus, the generation of mutants on a large scale is inevitable to understand the complete process or cycle. Several methods, including T-DNA insertion, TILLING (targeting-induced local lesions in genomes) and RNAi (RNA interference), were commonly used to create large-scale mutant libraries. Each of these methods has their own drawbacks like random insertions, difficulty to locate the inserted position and reduction in expression rather than the total knockout. Recent advances in genome engineering led to the development of precise genome editing methods, including CRISPR-Cas9, which has a high success rate in editing genome at the desired position, including the introduction of biallelic mutation. CRISPR/Cas9-based screening is target-specific and helps in identifying both functional genes and mutant phenotypes. CRISPR/Cas9-based robust methods would help to decode the functions of several hundred genes; in due course, it will be used to understand the genome function rather than the gene function.
