US 12,065,700 B2
Single sperm gene expression and mutation analysis for prediction of diseases
Christopher E. Mason, Ithaca, NY (US); Jeffrey Rosenfeld, Bergenfield, NJ (US); and Delia Tomoiaga, Ithaca, NY (US)
Assigned to CORNELL UNIVERSITY, Ithaca, NY (US); and Rutgers, The State University of New Jersey, New Brunswick, NJ (US)
Appl. No. 16/486,611
Filed by CORNELL UNIVERSITY, Ithaca, NY (US); and RUTGERS, THE STATE UNIVERSITY OF NEW JERSEY, New Brunswick, NJ (US)
PCT Filed Feb. 17, 2018, PCT No. PCT/US2018/018534
§ 371(c)(1), (2) Date Aug. 16, 2019,
PCT Pub. No. WO2018/152432, PCT Pub. Date Aug. 23, 2018.
Claims priority of provisional application 62/460,408, filed on Feb. 17, 2017.
Prior Publication US 2020/0010896 A1, Jan. 9, 2020
Int. Cl. C12Q 1/68 (2018.01); C12Q 1/6827 (2018.01); C12Q 1/6883 (2018.01); G16B 20/20 (2019.01); G16B 30/20 (2019.01); G16H 50/30 (2018.01)
CPC C12Q 1/6883 (2013.01) [C12Q 1/6827 (2013.01); G16B 20/20 (2019.02); G16B 30/20 (2019.02); G16H 50/30 (2018.01); C12Q 2600/156 (2013.01); C12Q 2600/158 (2013.01)] 21 Claims
 
1. A method comprising
isolating single sperm cells from a sperm sample;
constructing single cell RNA-Seq libraries, direct RNA measurement libraries, or cDNA libraries from the isolated single sperm cells;
generating a gene expression profile of each of MIR762HG, RP11-315I20.1, TADA2A, MEST, RP11-480I12.10, TMCO2, ZFP36L1, ATP1B3, CRAMP1, SPTY2D1-AS1, TNP1, AC007557.1, ACAP1, MARCKS, CTD-2568A17.1, PRM1, TCEB2, and TMEM31 based on the constructed single cell RNA-Seq libraries, direct RNA measurement libraries, or cDNA libraries;
comparing expression levels of the genes in the gene expression profile with reference expression levels of the same genes in a control healthy sperm sample;
determining that none of the genes in the expression profile are upregulated compared to the level of expression of the respective same gene in a healthy control sperm sample; and
utilizing the sperm sample for in vitro fertilization.