Targeting of the T-cell receptor zeta-chain gene in embryonic stem cells: strategies for generating multiple mutations in a single gene.

摘要: Abstract The T-cell receptor zeta chain is a member of family related proteins that play critical role in coupling cell-surface receptors to intracellular signaling pathways. To study the ontogeny, we generated targeted mutations zeta-chain gene murine embryonic stem cells. The mutant alleles are predicted result either null phenotype or synthesis truncated protein capable supporting T-cell-receptor surface expression but deficient transmembrane signaling. Both these targeting events were recovered single electroporation experiment with coelectroporation combination deletion/truncation construct. Our results suggest similar approaches could be used generate multiple mutations, modifications more than one site within gene, subtle alterations rely upon coconversion selectable marker gene.