US 12,454,690 B2
Type VI-E and type VI-F CRISPR-Cas system and uses thereof
Hui Yang, Shanghai (CN); Chunlong Xu, Shanghai (CN); Yingsi Zhou, Shanghai (CN); and Qingquan Xiao, Shanghai (CN)
Assigned to HuidaGene Therapeutics (Singapore) Pte. Ltd., Singapore (SG)
Filed by Huidagene Therapeutics (Singapore) Pte., Ltd., Singapore (SG)
Filed on Jan. 4, 2022, as Appl. No. 17/568,183.
Application 17/568,183 is a division of application No. 16/864,982, filed on May 1, 2020, granted, now 11,225,659.
Application 16/864,982 is a continuation of application No. PCT/CN2020/077211, filed on Feb. 28, 2020.
Prior Publication US 2022/0119808 A1, Apr. 21, 2022
Int. Cl. C12N 15/00 (2006.01); C12N 9/22 (2006.01); C12N 15/11 (2006.01); C12N 15/90 (2006.01)
CPC C12N 15/11 (2013.01) [C12N 9/22 (2013.01); C12N 15/907 (2013.01); C12N 2310/20 (2017.05); C12N 2800/80 (2013.01)] 26 Claims
 
1. A Clustered Regularly Interspaced Short Palindromic Repeat (CRISPR)-Cas system, comprising:
(1) an RNA guide sequence comprising a spacer sequence capable of hybridizing to a target RNA, and a direct repeat (DR) sequence 3′ to the spacer sequence, wherein the spacer sequence is between 25-50 nucleotides, wherein the DR sequence is an RNA sequence encoded by and corresponding to the full-length of the reverse complement sequence of SEQ ID NO: 8; and,
(2) a functional fragment of a CRISPR-associated protein (Cas), wherein said Cas has the amino acid sequence of SEQ ID NO: 1, wherein said functional fragment has an N-terminal deletion of no more than 210 residues, and a C-terminal deletion of no more than 180 residues, wherein the N-terminal deletion and C-terminal deletion result in the loss of RxxxxH motifs at both ends of the Cas having the amino acid sequence of SEQ ID NO: 1, wherein said functional fragment has completely or partially lost guide RNA-activated RNase catalytic activity compared to the guide RNA-activated RNase catalytic activity of said Cas having the amino acid sequence of SEQ ID NO: 1, and wherein said functional fragment is capable of (i) binding to the RNA guide sequence and (ii) targeting the target RNA.