Compositions for inducing modifications of target endogenous nucleic acid sequences in nucleuses of eukaryotic cells
AssigneeToolgen Incorporated
InventorsJin-Soo Kim, Seung Woo Cho, Sojung Kim
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Abstract
The present disclosure relates to targeted genome editing in eukaryotic cells or organisms. More particularly, the present disclosure provides for compositions that may induce modifications in target endogenous nucleic acid sequences in nucleuses of eukaryotic cells. The composition may comprise a single-chain guide RNA (sgRNA) and a Streptococcus pyogenes Cas9 protein. In some embodiments, the sgRNA and the Cas9 protein may be present in a molar ratio ranging from 29:14.0 to 29:1.4.
Claims13
A method of inducing a modification of a target endogenous nucleic acid sequence in a nucleus of a eukaryotic cell, comprising:
- preparing a single-chain guide RNA (sgRNA), wherein the sgRNA comprises a crRNA and a tracrRNA;
- preparing a Cas9 protein, wherein the Cas9 protein comprises a nuclear localization signal (NLS);
- combining the sgRNA and the Cas9 protein in vitro to form a Cas9/sgRNA complex, wherein the sgRNA and the Cas9 protein are present in a molar ratio ranging from 29:14 to 29:1.4;
- and
- introducing the Cas9/sgRNA complex into the eukaryotic cell, whereby the Cas9/sgRNA complex induces the modification of the target endogenous nucleic acid sequence in the nucleus of the eukaryotic cell.
Dependencies are inferred from the claim text.