Synthetic genes and genetic constructs

Patent metadata

Field	Value
Publication number	US-10006048-B2
Application number	US-201615199394-A
Country	US
Kind code	B2
Filing date	Jun 30, 2016
Priority date	Mar 20, 1998
Publication date	Jun 26, 2018
Grant date	Jun 26, 2018

Abstract

The present invention relates generally to synthetic genes for modifying endogenous gene expression in a cell, tissue or organ of a transgenic organism, in particular a transgenic animal or plant. More particularly, the present invention provides novel synthetic genes and genetic constructs which are capable of repressing delaying or otherwise reducing the expression of an endogenous gene or a target gene in an organism when introduced thereto.

First claim

The invention claimed is:

1. A double-stranded DNA construct comprising a first structural gene sequence comprising 20 consecutive nucleotides identical in sequence to a region of a target gene in an animal cell; a second structural gene sequence comprising 20 consecutive nucleotides identical in sequence to, and in an inverted orientation relative to, the 20 consecutive nucleotides of the first structural gene sequence, such that a repeating sequence which comprises the 20 consecutive nucleotides in length identical to the region of the target gene is present in the DNA construct; a stuffer fragment which consists of nucleotides other than the nucleotides of the repeating sequence and which separates and links the first and second structural gene sequences; a promoter and a transcription termination sequence, wherein the repeating sequence within the DNA construct comprises 20 nucleotides in length, and wherein the first structural gene sequence, the stuffer fragment and the second structural gene sequence are all operably connected to the promoter and the transcription termination sequence.

2. The double-stranded DNA construct of claim 1 , wherein the target gene is endogenous to the animal cell.

3. The double-stranded DNA construct of claim 1 , wherein the target gene is non-endogenous to the animal cell.

4. The double-stranded DNA construct of claim 1 , wherein the region of the target gene is in an exon.

5. The double-stranded DNA construct of claim 1 , wherein the target gene is from a lentivirus.

6. The double-stranded DNA construct of claim 1 , wherein the target gene is from a single-stranded (+) RNA virus.

7. The double-stranded DNA construct of claim 1 , wherein the animal cell is an insect cell.

8. An isolated animal cell, tissue or organ, having a DNA comprising a first structural gene sequence comprising 20 consecutive nucleotides identical in sequence to a region of a target gene in the animal cell; a second structural gene sequence comprising 20 consecutive nucleotides identical in sequence to, and in an inverted orientation relative to, the 20 consecutive nucleotides of the first structural gene sequence, such that a repeating sequence which comprises the 20 consecutive nucleotides in length identical to the region of the target gene is present in the DNA, a stuffer fragment which consists of nucleotides other than the nucleotides of the repeating sequence and which is between and links the first and second structural gene sequences; a promoter and a transcription termination sequence, wherein the repeating sequence within the DNA comprises 20 nucleotides in length, and wherein the first structural gene sequence, the stuffer fragment and the second structural gene sequence are all operably connected to the promoter and the transcription termination sequence.

9. The isolated animal cell of claim 8 , wherein the target gene is endogenous to the animal cell.

10. The isolated animal cell of claim 8 , wherein the target gene is non-endogenous to the animal cell.

11. The isolated animal cell, tissue or organ of claim 8 , wherein the region of the target gene is in an exon.

12. The isolated animal cell, tissue or organ of claim 8 , wherein the target gene is from a lentivirus.

13. The isolated animal cell, tissue or organ of claim 8 , wherein the target gene is from a single-stranded (+) RNA virus.

14. The isolated animal cell, tissue or organ of claim 8 , wherein the DNA is integrated into the genome of the isolated animal cell, tissue or organ.

15. The isolated animal cell of claim 8 , wherein the animal cell is an insect cell.

Assignees

• Commw Scient Ind Res Org

Inventors

• Graham Michael Wayne
• Rice Robert Norman

Classifications

• C12N2830/38

  being a stuffer · CPC title

• C12N9/0071

  acting on paired donors with incorporation of molecular oxygen (1.14) · CPC title

• C12N9/503

  derived from viruses · CPC title

• C12N2840/102

  inhibiting translation · CPC title

• C12N2830/00

  Vector systems having a special element relevant for transcription · CPC title