Patent ID: 8680256
Filing Date: 2014-03-25
Classification: C12N

Abstract:
1. A method for producing inducible expression active linear gene constructs, comprising the step of: (a) generating an inducible expression active linear gene construct, comprising wherein the generating step comprises a PCR amplification of a source expression polynucleotide comprising in 5′ to 3′ direction a promoter sequence and the DNA sequence using wherein the source expression polynucleotide is a vector, a lentivirus, a plasmid, a virus-based vector, or a linear or linearized or amplified fragment thereof, wherein the mRNA destabilization elements are selected from AU-rich elements, GU-rich elements, and U-rich sequences, wherein the mRNA stabilization elements are selected from GC-rich elements, CU-rich elements, and UG-rich sequences, wherein the generating step does not involve a cloning step, wherein said cloning step includes the use of one or more of restriction digestion, cloning enzyme(s), bacterial transformation and plasmid preparation, and wherein expression of the inducible expression active linear gene construct resulting from (a) can be induced by the addition of a compound that activates the expression or by the withdrawal of a compound that represses the expression.