公开(公告)号：US11834670B2

公开(公告)日：2023-12-05

申请号：US15491125

申请日：2017-04-19

申请人： Global Life Sciences Solutions USA LLC

发明人： John Richard Nelson ,  Robert Scott Duthie ,  Patrick McCoy Spooner ,  John Anthony Schiel ,  Lisa Anne Lowery ,  Anja Josifa Smith

IPC分类号： C12N15/11 ,  C12N15/90 ,  C12N15/10

CPC分类号： C12N15/907 ,  C12N15/102 ,  C12N15/11 ,  C12N2310/20

摘要： A method of site-specific modification of an endogenous target DNA of a eukaryotic cell is provided. The method includes contacting the endogenous target DNA having an intended modification site with (i) a gene editing system configured to introduce a double strand break in the endogenous target DNA at or near the intended modification site, and (ii) a donor DNA repair template comprising a plurality of tandem repeat sequences. In the method, each of the plurality of tandem repeat sequences comprises an exogenous donor DNA sequence flanked by a donor 5′ flanking sequence and a donor 3′ flanking sequence. The donor 5′ flanking sequence and the donor 3′ flanking sequence are homologous to a continuous DNA sequence on either side of the intended modification site in the endogenous target DNA.

公开(公告)号：US11987836B2

公开(公告)日：2024-05-21

申请号：US15510331

申请日：2015-09-17

申请人： Global Life Sciences Solutions USA LLC

发明人： Ryan Charles Heller ,  Nichole Lea Wood ,  Robert Scott Duthie ,  John Richard Nelson ,  Wei Gao ,  Michael James Rishel ,  Klaus Gustav Hentrich

IPC分类号： C12Q1/6806 ,  C12Q1/6844 ,  C12Q1/6858 ,  C12Q1/6869

CPC分类号： C12Q1/6806 ,  C12Q1/6844 ,  C12Q1/6858 ,  C12Q1/6869 ,  C12Q1/6844 ,  C12Q2521/501 ,  C12Q2531/125

摘要： A method is provided for generating single-stranded DNA circles from a biological sample. The method comprises the steps of: treating the biological sample with an extractant to release nucleic acids, thereby forming a sample mixture; neutralizing the extractant; denaturing the released nucleic acids to generate single-stranded nucleic acids; and contacting the single-stranded nucleic acids with a ligase that is capable of template-independent, intramolecular ligation of single-stranded DNA to generate the single-stranded DNA circles. All the steps of the method are performed without any intermediate nucleic acid isolation or nucleic acid purification. The single-stranded DNA circles may be amplified and further analyzed. Also provided is a kit which comprises compositions for carrying out the novel methods.