	US 12,213,468 B2
	Population control using engineered translocations
Bruce A. Hay, Encino, CA (US); Omar S. Akbari, Pasadena, CA (US); and Anna B. Buchman, Pasadena, CA (US)
Assigned to California Institute of Technology, Pasadena, CA (US)
Filed by California Institute of Technology, Pasadena, CA (US)
Filed on Aug. 27, 2020, as Appl. No. 17/004,698.
Application 17/004,698 is a division of application No. 15/164,452, filed on May 25, 2016, granted, now 10,966,414.
Claims priority of provisional application 62/166,281, filed on May 26, 2015.
Prior Publication US 2020/0404892 A1, Dec. 31, 2020
Int. Cl. A01K 67/033 (2006.01); C12N 15/85 (2006.01)

CPC A01K 67/0339 (2013.01) [C12N 15/8509 (2013.01); A01K 2217/15 (2013.01); A01K 2227/706 (2013.01); A01K 2267/02 (2013.01)]

14 Claims

1. A method of distributing one or more genes of interest into a population of insects, the method comprising:

i) providing an insect population, wherein one or more individuals in the insect population comprises a translocation mediated gene drive system; and

ii) inducing a chromosomal translocation in the one or more individuals in the insect population,

wherein the chromosomal translocation generates a translocation-bearing altered insect population, wherein the translocation-bearing altered insect population comprises translocation-bearing individuals that are translocation heterozygotes and translocation-bearing individuals that are translocation homozygotes for the chromosomal translocation and wherein the translocation mediated gene drive system comprises:

a first construct, configured to be positionable at a first insertion site in a first chromosome, wherein the first construct comprises:

a) a first location to insert a first gene of interest;

b) a first promoter;

c) a first fragment of foreign stuffer DNA;

d) a second fragment of foreign stuffer DNA;

e) a first target site of a pair of target sites for an endonuclease positioned between the first and second fragments of foreign stuffer DNA;

f) a first splice acceptor site, positioned downstream from a-e; and

g) a first splice donor site, positioned between b and c, and a second construct, configured to be positionable at a second insertion site in a second chromosome, wherein the second construct comprises:

h) a second location to insert a second gene of interest;

i) a second promoter;

j) a third fragment of foreign stuffer DNA, wherein the third fragment is homologous to the second fragment;

k) a fourth fragment of foreign stuffer DNA, wherein the fourth fragment is homologous to the first fragment;

l) a second target site of the pair of target sites for an endonuclease positioned between the third and fourth fragments of foreign stuffer DNA;

m) a second splice acceptor site, positioned downstream from h-1, and

n) a second splice donor site, positioned between i and j, wherein the first and second chromosomes are non-homologous chromosomes,

wherein the first fragment of foreign stuffer DNA is homologous to the fourth fragment of foreign stuffer DNA and the second fragment of foreign stuffer DNA is homologous to the third fragment of foreign stuffer DNA,

wherein a double stranded break created at the first, and second target sites of the pair of target sites allows for homologous recombination between the first and fourth fragments, and between the second and third fragments upon a repair of the double stranded break (DSB),

wherein the repair of the DSB induces a chromosomal translocation and generates a first translocation chromosome and a second translocation chromosome.