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1. (WO2015048429) ENGINEERING DNA ASSEMBLY IN VIVO AND METHODS OF MAKING AND USING THE REVERSE TRANSCRIPTASE TECHNOLOGY

Pub. No.:    WO/2015/048429    International Application No.:    PCT/US2014/057695
Publication Date: Fri Apr 03 01:59:59 CEST 2015 International Filing Date: Sat Sep 27 01:59:59 CEST 2014
IPC: C12N 15/10
C12N 15/63
C12N 5/10
A61K 48/00
B82Y 40/00
B82Y 5/00
Applicants: MASSACHUSETTS INSTITUTE OF TECHNOLOGY
Inventors: ELBAZ, Johann
VOIGT, Christopher
Title: ENGINEERING DNA ASSEMBLY IN VIVO AND METHODS OF MAKING AND USING THE REVERSE TRANSCRIPTASE TECHNOLOGY
Abstract:
Cells that can synthesize oligonucleotides in vivo to produce a nucleic acid nanostmcture are described. Methods for producing oligonucleotide nano structures for use in regulating gene expression and altering biological pathways are provided. Methods of performing multiplex automated genome editing (MAGE) are also provided. Methods of synthesizing single stranded DNA (ssDNA) comprising expressing a reverse transcriptase in a cell and a functional template comprising a non-coding tRNA structure at the 3' end and a coding RNA sequence at the 5' end, wherein the non-coding tRNA structure is capable of initiating transcription of the coding RNA sequence using the reverse transcriptase to produce the ssDNA oligonucleotide in the cell are also provided.