Golden gate method in biology and medicine

The aim of this review was to describe and compare molecular cloning methods for assembling genetic constructs. Genetic therapy is one of the rapidly developing fields of modern medicine, so special attention in this review is paid to parameters such as speed, accuracy, and efficiency of cloning, as these are critical factors in creating gene therapy agents. Special attention is given to the Golden Gate method, which is based on the use of type IIS restriction endonucleases, as this approach simplifies the cloning process and increases its efficiency through standardized design and a minimal set of enzymes. Alongside Golden Gate, the review also discusses more innovative molecular cloning methods, Gateway and Gibson, in terms of their potential use for addressing fundamental and applied challenges in regenerative medicine.