US 12,234,450 B2
Generation of synthetic genomes
Clyde A. Hutchison, La Jolla, CA (US); Ray-Yuan Chuang, Rockville, MD (US); Vladimir N. Noskov, Gaithersburg, MD (US); Bogumil J. Karas, London (CA); Kim S. Wise, San Diego, CA (US); Hamilton O. Smith, San Diego, CA (US); John I. Glass, San Diego, CA (US); Chuck Merryman, Encinitas, CA (US); Daniel G. Gibson, Carlsbad, CA (US); J. Craig Venter, La Jolla, CA (US); Krishna Kannan, San Diego, CA (US); and Lin Ding, San Diego, CA (US)
Filed by Codex DNA, Inc., San Diego, CA (US)
Filed on Mar. 12, 2021, as Appl. No. 17/200,633.
Application 17/200,633 is a continuation of application No. 15/466,675, filed on Mar. 22, 2017, granted, now 11,085,037.
Claims priority of provisional application 62/312,398, filed on Mar. 23, 2016.
Prior Publication US 2021/0254046 A1, Aug. 19, 2021
This patent is subject to a terminal disclaimer.
Int. Cl. C12N 15/10 (2006.01); C12N 15/74 (2006.01); C12N 15/81 (2006.01); G16B 30/00 (2019.01); G16B 30/20 (2019.01)
CPC C12N 15/1027 (2013.01) [C12N 15/1058 (2013.01); C12N 15/1079 (2013.01); C12N 15/1093 (2013.01); C12N 15/74 (2013.01); C12N 15/81 (2013.01); G16B 30/00 (2019.02); G16B 30/20 (2019.02)] 20 Claims
 
1. A method for generating a synthetic genome of interest, comprising:
(a) providing a first genome sufficient to sustain viability of a prokaryotic cell;
(b) designing and providing a second genome comprising a reduced number of genes compared to the first genome;
(c) dividing each of the first and second genomes into at least three corresponding fragments;
(d) combining at least one fragment of the second genome with fragments of the first genome to generate a third genome having all the at least three corresponding fragments, further comprising grouping genes related to the same biological process in at least one fragment of the second genome prior to combining; or grouping genes related to the same biological process in the third genome after combining;
(e) testing the third genome generated in step (d) for sufficiency to sustain viability of a cell; and
(f) identifying the third genome as a synthetic genome of interest if it sustains viability of the prokaryotic cell; otherwise genetically modifying the at least one fragment of the second genome and repeating steps (d)-(f) in one or more iterations until a genome that sustains viability of the prokaryotic cell is obtained in the third genome.