hnRNP A1 KNOCKOUT ANIMAL MODEL AND USE THEREOF

Abstract

A nucleic acid construct comprising a genetic engineered heterogeneous nuclear ribonucleoprotein (hnRNP) A1 gene is provided. A transgenic mouse in which the expression of hnRNP A1 gene has been disrupted is also provided. The mouse is useful for studying the role of hnRNP A1 gene in normal and disease states of a neurodegenerative disease or a cancer for developing therapies to treat any of these diseases. Therefore, a method of screening a compound for potential use in prevention and/or treatment of neurodegenerative disease or cancer is further provided.

1 . A nucleic acid construct comprising a genetic engineered heterogeneous nuclear ribonucleoprotein (hnRNP) A1 gene, wherein a first locus of recombination sequence 1 is inserted before exon 2 of the hnRNP A1 gene, and recombination sequences 2 flanking a marker gene followed by a second locus of recombination sequence 1 are inserted behind exon 8 of the hnRNP A1 gene;
 wherein the recombination sequence 1 and recombination sequence 2 undergo recombination with the same recombination sequence at the other locus in the presence of their corresponding recombinase.   
     
     
         2 . The nucleic acid construct of  claim 1 , wherein the marker gene detects the presence of the nucleic acid construct in a cell. 
     
     
         3 . The nucleic acid construct of  claim 1 , wherein the marker gene is an antibiotic resistance gene, a reporter lacZ gene, or a herpes simplex virus thymidine kinase gene (HSV-tk). 
     
     
         4 . The nucleic acid construct of  claim 3 , wherein the antibiotic resistance gene is neomycin-resistant gene. 
     
     
         5 . The nucleic acid construct of  claim 1 , wherein the recombination sequence 1 is a crossover (x) in P1 (loxP) site and the recombination sequences 2 is a FLP recombinase target (frt) site. 
     
     
         6 . The nucleic acid construct of  claim 5 , wherein the hnRNP A1 gene is disrupted by introduction of Cre recombinase and FLP recombinase. 
     
     
         7 . A transgenic mouse whose genome comprising a homozygous disruption in hnRNP A1 gene, wherein the disruption results in the transgenic mouse having a damaged function of exonic RNA splicing, a reduced weight relative to a wild-type control mouse at embryonic stage, and perinatal mortality. 
     
     
         8 . The transgenic mouse of  claim 7 , wherein the disruption affects at least two exons within the hnRNP A1 gene. 
     
     
         9 . The transgenic mouse of  claim 8 , wherein the exons are exons 2 to 8. 
     
     
         10 . A transgenic mouse whose genome comprising a heterozygous disruption in hnRNP A1 gene. 
     
     
         11 . The transgenic mouse of  claim 10 , wherein the disruption affects at least two exons within the hnRNP A1 gene. 
     
     
         12 . The transgenic mouse of  claim 11 , wherein the exons are exons 2 to 8. 
     
     
         13 . The transgenic mouse of  claim 10 , wherein the transgenic mouse is predisposed to premature aging diseases and/or virus infective diseases. 
     
     
         14 . A method of screening a compound for potential use in prevention and/or treatment of neurodegenerative disease or cancer, which comprises,
 (a) administering a test compound to the transgenic mouse of  claim 7  or primary cells or tissues derived therefrom;   (b) administering a test compound to a wild-type mouse or primary cells or tissues derived therefrom;   (c) assessing functions of exonic RNA splicing in each of the mice, cell, or tissues from step (a) and (b), prior to and after a given time period of the administration; and   (d) comparing the assessment results of step (c) to determine effectiveness of the test compound.