A study was undertaken for genetic diversity analysis of three native sheep populations of Eastern Indian Himalaya utilizing twenty five ovine microsatellite markers. A total of 150 sheep, 50 each from the states of Assam, Meghalaya and Arunachal Pradesh were genotyped through microsatellite. Polymorphism was revealed in most of the markers with a total of 242 alleles. The average observed (Ho) and expected (He) heterozygosity estimates for Assam, Meghalaya and Arunachal Pradesh sheep were 0.311 and 0.396; 0.323 and 0.383; and 0.416 and 0.408, respectively suggesting that the genetic diversity of each population ranged from intermediate to high. The average polymorphism information content (PIC) estimates and Shannon’s Information index were 0.357 and 0.70, 0.366 and 0.75, and 0.292 and 0.75 in Assam, Meghalaya and Arunachal Pradesh sheep populations. Mean FIS (within- population inbreeding estimate), FIT (Between-population inbreeding estimate) and FST (Population differentiation measure) estimate for three populations under consideration were 0.149, 0.224 and 0.088, respectively. Significant deviations from Hardy-Weinberg equilibrium (based on chi-square) were found in a total of 10, 13 and 6 alleles in Assam, Meghalaya and Arunachal Pradesh sheep population which indicated evolutionary force in operation at these loci in the respective population. The study revealed that Assam sheep and Meghalaya sheep are the closest relatives whereas Assam sheep and Arunchal Pradesh sheep are farthest relatives.