Cholesterol biosynthesis is an essential, tightly controlled biochemical process in human physiology. It is an essential part of membrane fluidity, cellular homeostasis, and for synthesis of steroid hormone and bile acid. The review will address the fine balance of enzymatic and genetic modulators governing endogenous cholesterol synthesis and emphasis will be laid on the importance and regulation. Mevalonate is produced first upon acetyl-CoA conversion. It is regulated by HMGCR enzyme that is also referred to as the rate limiting enzyme. Cholesterol is then biosynthesized through consecutive enzymatic conversions, with rate limiting enzymes including squalene epoxidase (SQLE), squalene synthase (FDFT1) and 7-dehydrocholesterol reductase (DHCR7). These enzymes are rigorously regulated by sterol regulatory element-binding proteins (SREBPs) through a complicated feedback mechanism that maintains metabolic balance. However, due to the multiple reactions involved, any dysfunction or disruptions in the expression of these regulatory genes, a variety disorders may be developed, comprising hyperlipidemia, atherosclerosis, various malignancies, and rare inherited enzymopathies, for instance, Smith-Lemli-Opitz syndrome (SLOS). From the clinical standpoint, various therapies have emerged to modulate major elements of the pathway, such as statins, PCSK9 inhibitors, and drugs to modulate SREBP signaling. These remain key to the management of cholesterol disorders and the betterment of patient outcomes