US 12,442,004 B2
Linear covalently closed vectors and related compositions and methods thereof
Nikolai Eroshenko, Boston, MA (US); and Nikhil Dhar, Boston, MA (US)
Assigned to Helix Nanotechnologies Inc, Boston, MA (US)
Filed by Helix Nanotechnologies Inc, Boston, MA (US)
Filed on Aug. 29, 2022, as Appl. No. 17/897,498.
Application 17/897,498 is a division of application No. 16/614,101, granted, now 11,473,090, previously published as PCT/US2018/032988, filed on May 16, 2018.
Claims priority of provisional application 62/507,144, filed on May 16, 2017.
Prior Publication US 2023/0193293 A1, Jun. 22, 2023
Int. Cl. C12Q 1/68 (2018.01); C12N 15/63 (2006.01); C12N 15/66 (2006.01); C12Q 1/6844 (2018.01); C12Q 1/6897 (2018.01)
CPC C12N 15/66 (2013.01) [C12N 15/63 (2013.01); C12Q 1/6846 (2013.01); C12Q 1/6897 (2013.01)] 15 Claims
OG exemplary drawing
 
1. A composition comprising:
(i) a first payload oligonucleotide including a first payload sequence and a first 5′ extension sequence;
(ii) a second payload oligonucleotide including a second payload sequence and a second 5′ extension sequence, wherein the first payload sequence is complementary to the second payload sequence;
(iii) a first adaptor oligonucleotide comprising:
(a) a first stem sequence and a second stem sequence, wherein the first stem sequence is an inverted complement of the second stem sequence, and
(b) a first adaptor 5′ extension sequence that is complementary to the first 5′ extension sequence;
(iv) a second adaptor oligonucleotide comprising:
(a) a third stem sequence and a fourth stem sequence, wherein the third stem sequence is an inverted complement of the fourth stem sequence, and
(b) a second adaptor 5′ extension sequence that is complementary to the second 5′ extension sequence;
(v) a third payload oligonucleotide including a third payload sequence and a first 3′ extension sequence;
(vi) a fourth payload oligonucleotide including a fourth payload sequence and a second 3′ extension sequence, wherein the third payload sequence is complementary to the fourth payload sequence;
(vii) a third adaptor oligonucleotide comprising:
(a) a fifth stem sequence and a sixth stem sequence, wherein the fifth stem sequence is an inverted complement of the sixth stem sequence, and
(b) a first adaptor 3′ extension sequence that is complementary to the first 3′ extension sequence; and
(viii) a fourth adaptor oligonucleotide comprising:
(a) a seventh stem sequence and an eighth stem sequence, wherein the seventh stem sequence is an inverted complement of the eighth stem sequence, and
(b) a second adaptor 3′ extension sequence that is complementary to the second 3′ extension sequence.