Methods and compositions for increasing transgene activity
Methods and compositions for increasing transgene expression and/or activity, including for increasing nuclease-mediated genomic modifications.
1. A method of increasing nuclease activity in a cell, the method comprising: introducing an mRNA comprising a 3′ untranslated region (3′ UTR) and encoding at least one component of one or more nucleases into the cell, wherein the mRNA comprises a woodchuck hepatitis virus posttranslational regulatory element (WPRE) as shown in any of SEQ ID NOs:1-3 in the 3′ UTR.
2. The method of claim 1 , wherein the WPRE is a wild-type WPRE.
3. The method of claim 1 , wherein the nuclease is a zinc finger nuclease, a TALEN and/or a CRISPR/Cas and/or a Cfp1 CRISPR/Cas nuclease system.
4. The method of claim 1 , further comprising subject the cells to cold-shock conditions.
5. An isolated mRNA comprising a WPRE sequence as shown in any of SEQ ID NO:1-3 in the 3′ UTR and a sequence encoding at least one component of a nuclease or a transcription factor (TF).
6. The mRNA of claim 5 , wherein the sequence encodes a nuclease.
7. The mRNA of claim 5 , wherein the nuclease is a zinc finger nuclease, a TALEN and/or at least one component of a CRISPR/Cas nuclease system.
8. The mRNA of claim 5 , wherein the Cas protein is a Cas and/or Cfp1 protein.
9. The mRNA of claim 5 , wherein the sequence encodes a transcription factor.
10. The mRNA of claim 9 , wherein the transcription factor is a ZF-TF, a TALE-TF or a CRISPR/Cas TF.
11. The mRNA of claim 5 , further comprising a polyA sequence.