32 RRJBS| Volume 4 | Issue 2 | August-September, 2015 Research & Reviews: Journal of Botanical Sciences e-ISSN:2320-0189 p-ISSN:2347-2308 INTRODUCTION Cannabis (Cannabis sativa L., Cannabaceae) is an annual herbaceous plant in the Cannabis genus, indigenous to Central and South Asia. Cannabis produces many secondary compounds such as lavonoids, stilbenoids, alkaloids, lignanamides, phenolic amides, but main active psychotropic constituent of cannabis is Δ 9 -tetrahydrocannabinol (Δ 9 -THC). Cannabinoids, terpenoids, and other compounds are secreted by glandular trichomes that occur most abundantly on the loral calyxes and bracts of female plant. Despite of its medicinal properties, C. sativa is among the very oldest of economic plants providing humans with iber for spinning, weaving cloth, and making paper; seed for human foods and animal feeds; and aromatic resin containing compounds of recreational and medicinal value and also a useful source of foodstuffs and bio-fuels [1,2] . The recent progress in functional genomics research based on large-scale expressed sequence tag (EST) generation, analysis and cloning of genes in cannabis plant has provided a critical signiicance on elucidating the molecular mechanism of growth, development, differentiation, metabolism, quality, yield, and stress resistance, as well as genetic manipulation via biotechnological approaches in the foreseeable future. However, in cannabis, only a few regulatory players mostly related to regulating gland development and THCA biosynthesis have been studied so far [3,4] . Up to date, limited progress has been made Computational Prediction, Target Identification and Experimental Validation of miRNAs from Expressed Sequence Tags in Cannabis sativa . L Ganesh Selvaraj Duraisamy 1 , Ajay Kumar Mishra 1 , Jernej Jakse 2 and Jaroslav Matousek 1* 1 Biology Centre ASCR v.v.i, Institute of Plant Molecular Biology, Branisovska 31, ceske Budejovice 370 05, Czech Republic 2 University of Ljubljana, Biotechnical Faculty, Agronomy Department, Jamnikarjeva 101, SI-1000 Ljubljana, Slovenia Research Article ABSTRACT MicroRNAs (miRNAs) are approximately 20-22 nucleotide non-coding RNAs, which play an important role in posttranscriptional degradation of target mRNA or inhibition of protein synthesis through binding the speciic sites of target mRNA. miRNAs have been extensively studied in various plant species, however, there is no miRNA identiied in Cannabis sativa, a crop which has long been used for hemp ibre, for seed and seed oils, for medicinal purposes, and as a recreational drug. In this study, a computational based approach was used to identify and characterize C. sativa miRNAs. A total of 7 miRNAs belonging to 3 miRNA families were identiied in cannabis based on homolog search and series of iltering criteria. The identiied miRNAs were validated by end- point PCR and quantitative reverse transcription-polymerase chain reaction (qRT-PCR), conirmed the existence of conserved miRNAs in C. sativa. Based on near-perfect complementarity between cannabis miRNAs and their target mRNA gene sequence, a total of 23 miRNAs targets were identiied involved in processes such as plant development, signal transduction, secondary metabolite production, protein degradation, response to environmental stress and pathogen invasion, and ability to regulate their own biogenesis. The cis- regulatory elements relevant to biotic and abiotic stress, plant hormone response lavonoid and cannabinoids biosynthesis were identiied in the promoter regions of those miRNA genes. Overall, indings from this study will accelerate the way for further researches of miRNAs and their functions in C. sativa, particularly cannabinoid metabolic pathway. Received date: 21/08/2015 Accepted date: 09/09/2015 Published date: 11/09/2015 *For Correspondence Jaroslav Matousek , Biology Centre ASCR v.v.i, Institute of Plant Molecular Biology, Branisovska 31, ceske Budejovice 370 05, Czech Republic ,Tel: (+420) 387 775 525 E-mail: jmat@umbr.cas.cz Keywords: Cannabis sativa, microRNAs, Cis-regulating elements, Computational approach, EST, Targets