The invention discloses shRNA for inhibiting human
EDARADD gene expression, a lentiviral vector and a construction method and application of the lentiviral vector. The method comprises the following steps: firstly, synthesizing a positive strand and a
negative strand of
DNA oligo aiming at an
RNA interference target sequence; annealing the positive strand and the
negative strand to form double-stranded
DNA with a cohesive end; linearizing the lentiviral vector through
enzyme digestion, and connecting the linearized vector with double-stranded
DNA; transforming
escherichia coli competent cellswith a connection product, and carrying out PCR identification on the obtained positive clone; and performing plasmids extraction for
virus packaging in the next step. According to the invention, theRNA interference target sequence is designed by taking an
EDARADD gene as a template, an shRNA interference sequence is designed according to a selected target sequence, an
RNA interference lentiviralvector is constructed through packaging, and after the
EDARADD gene is knocked down, proliferation of
tongue squamous cell carcinoma cells CAL-27 can be remarkably influenced, the
cell clone formingcapacity is influenced, and
cell apoptosis is promoted.