Modified cells comprising CRISPR systems
AssigneeThe Regents of the University of California
InventorsJennifer A. Doudna, Basem Al-Shayeb, Jillian F. Banfield, Patrick Pausch
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Abstract
The present disclosure provides RNA-guided CRISPR-Cas effector proteins, nucleic acids encoding same, and compositions comprising same. The present disclosure provides ribonucleoprotein complexes comprising: an RNA-guided CRISPR-Cas effector protein of the present disclosure; and a guide RNA. The present disclosure provides methods of modifying a target nucleic acid, using an RNA-guided CRISPR-Cas effector protein of the present disclosure and a guide RNA. The present disclosure provides methods of modulating transcription of a target nucleic acid.
Claims32
A eukaryotic cell comprising:
- a) a nuclease comprising an amino acid sequence that is at least 98% identical to the amino acid sequence set forth in SEQ ID NO:120;
- b) a target nucleic acid comprising a target strand and a non-target strand;
- and
- c) a recombinant guide nucleic acid comprising in a 5′ to 3′ orientation:
- i) a first region that binds to the nuclease;
- and
- ii) a second region that is complementary to a portion of the target strand of the target nucleic acid,
- wherein the target nucleic acid comprises a protospacer adjacent motif (PAM) of 5′-NTTN-3′, wherein T is thymine and N is any nucleotide,
- and wherein the PAM is on the non-target strand of the target nucleic acid.
A eukaryotic cell comprising
- A) a target nucleic acid comprising a target strand and a non-target strand,
- wherein the target nucleic acid comprises a protospacer adjacent motif (PAM) of 5′-NTTN-3′, wherein T is thymine and N is any nucleotide,
- and wherein the PAM is on the non-target strand of the target nucleic acid;
- and
- B) at least one nucleic acid molecule encoding:
- a) a nuclease comprising an amino acid sequence that is at least 98% identical to the amino acid sequence set forth in SEQ ID NO:120;
- and
- b) a recombinant guide nucleic acid comprising in a 5′ to 3′ orientation:
- i) a first region that binds to the nuclease;
- and
- ii) a second region that is complementary to a portion of the target strand of the target nucleic acid.
Dependencies are inferred from the claim text.