The invention discloses a method for constructing a
genetic engineering cell strain of an
obesity-resistant
medicine target point UCP1, and besides, discloses establishing and application of an
obesity-resistant
medicine high-flux screening model. Mainly a
CRISPR / Cas9
system is related and utilized, two unique sgRNAs are designed,
luciferase-T2A-tdTomato-WPRE-pA is knocked in after N end ATG of aUCP1
gene of cells, particularly
luciferase and tdTomato are knocked in
gene sites of immortalization brown fat cells UCP1, a first stably-transfected brown fat
cell strain in which
luciferase and tdTomato are inserted in a
promoter region of the UCP1 is formed, and the first stably-transfected brown fat
cell strain is applied to high-flux screening of
obesity-resistant medicines, evaluation of the obesity-resistant medicines, evaluation of obesity-resistant active substances of organisms and development of a UCP1 detection
reagent kit. The UCP1 is
uncoupling protein specifically expressed atbrown fat tissue, and is an obesity-resistant new target point. The construction of the stably-transfected
genetic engineering cell strain has important significance in the aspects of applying reportgenes and a high-connotation method, performing high-flux screening on a compound
library acutely, accurately and efficiently, and obtaining obesity-resistant medicines capable of promoting
thermogenesis and reducing weight.