ISOLATION OF LIVING CELLS AND PREPARATION OF CELL LINES BASED ON DETECTION AND QUANTIFICATION OF PRESELECTED CELLULAR RIBONUCLEIC ACID SEQUENCES

摘要

The invention is directed to reliable and efficient detection of mRNAs as well as other RNAs in living cells and its use to identify and, if desired, separate cells based on their desired characteristics. Such methods greatly simplify and reduce the time necessary to carry out previously-known procedures, and offers new approaches as well, such as selecting cells that generate a particular protein or antisense oligonucleotide, generating cell lines that express multiple proteins, generating cell lines with knock-out of one or more protein, and others.

主权项

1. A method for generating a cell line expressing at least one preselected protein comprising the steps of:
a) transfecting cell lines with at least one DNA construct encoding said at least one preselected protein and at least one drug resistance marker; b) selecting for cells resistant to a drug to which said marker confers resistance, said cells transcribing the at least one DNA construct and the at least one drug resistance marker; c) exposing said selected cells to a first molecular beacon that fluoresces upon hybridization to an RNA transcript of said at least one preselected protein; d) isolating said cells that fluoresces; e) generating a cell line expressing said at least one preselected protein by growing said isolated cells.