Patent ID: 8716021

Claim:
An ex vivo method of inhibiting silencing of a gene in a cell, comprising: (A) introducing into the cell a transgene construct comprising: (i) a coding nucleic acid sequence to be expressed in the cell; and (ii) a metazoan replicator nucleic acid sequence, wherein the metazoan replicator nucleic acid sequence integrates into a host genome with the coding nucleic acid sequence and: (a) is between 1.2 kb and 5.8 kb in length; (b) comprises a nucleic acid sequence at least 95% identical to the nucleic acid sequence set forth as SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, SEQ ID NO: 4, SEQ ID NO: 5, SEQ ID NO: 6, or SEQ ID NO: 7; (c) confers initiation of DNA replication in cis at ectopic chromosomal sites; (d) maintains DNA replication of the transgene construct in early S phase; (e) inhibits chromatin condensation; and (f) inhibits silencing of expression of the coding nucleic acid sequence; and (B) detecting expression of the coding nucleic acid sequence at least six weeks after introducing the metazoan replicator nucleic acid sequence into the cell, compared to a control cell that does not comprise the replicator nucleic acid sequence, thereby inhibiting silencing of the gene in the cell.