12 results about "Sequence Insertions" patented technology

Provided is an efficient and precise plant gene knockout method. First, a sequence containing a stop codon cluster is inserted into a genome by means of dual pegRNA to obtain, by means of screening, a stop codon cluster sequence capable of efficiently and precisely knocking out a target gene. A prime editing system and multiple pegRNA programming can be used in combination with the method to enable various genomic modifications such as efficient and precise knockout of one or more target genes, base substitutions, and small fragment insertions, deletions, and substitutions with just one prime editing protein.

The application discloses a method for identifying large fragment sequence insertion of a target genomic region and application thereof, and belongs to the technical field of bioinformatics. In view of the problem that large fragment insertion variation is difficult to be accurately recognized in clinical metagenomic sequencing due to short sequencing read length, insufficient coverage and other factors, the application proposes to construct a reference sequence which can represent the insertion variation by means of manual construction, and to realize efficient identification of the insertion event of the target genomic region by combining a short read-based fast alignment process. The method overcomes the dependence of existing structural variation detection tools on high sequencing depth and long read length, has the advantages of fast identification speed, high sensitivity and high accuracy, and is suitable for rapid screening of large fragment insertion related to drug resistance mechanism in clinical samples. Meanwhile, the method can be popularized for insertion variation analysis of other pathogen drug resistance related genes or genomic regions, and has a good clinical application prospect.

The present invention provides, among other things, methods of engineering a re-editable target locus in a cell, comprising inserting an exogenous re-editable template sequence to a target locus and methods of engineering a re-editable cell comprising a re-editable template sequence flanked by two homology arms such that the re-editable template sequence is inserted into a target locus by homologous recombination, and methods of using the same. In some aspects, provided is a re-editable template sequence, wherein the template sequence has no substantial sequence similarity to any region in the genome, and wherein the template sequence is recognizable by a genome editing system.

The invention provides, inter alia, a method of engineering a reeditable target locus in a cell, the method comprising inserting an exogenous reeditable template sequence into the target locus; a method of engineering a reeditable cell, the cell comprising a reeditable template sequence flanked by two homologous arms such that the reeditable template sequence is inserted into a target locus by homologous recombination, and a method of engineering a reeditable cell comprising a reeditable template sequence flanked by two homologous arms, and methods of use thereof. In some aspects, a reeditable template sequence is provided, where the template sequence has no substantial sequence similarity to any region in the genome, and where the template sequence can be identified by a genome editing system.

The invention discloses a method for identifying large-fragment sequence insertion in a target genome region and application of the method, and belongs to the technical field of bioinformatics. In order to solve the problem that large-fragment insertion variation is difficult to accurately identify due to factors such as short sequencing reading length and insufficient coverage in clinical metagenome sequencing, the invention proposes that a reference sequence capable of representing the insertion variation is artificially constructed and is combined with a rapid comparison process based on short reads to realize efficient identification of an insertion event in a target genome region. The method overcomes the dependence of an existing structure variation detection tool on high sequencing depth and long reading length, has the advantages of high identification speed, high sensitivity, high accuracy and the like, and is suitable for rapid screening of large fragment insertion related to a drug resistance mechanism in a clinical sample; meanwhile, the method can be popularized and applied to insertion variation analysis of other pathogen drug resistance related genes or genome areas, and has a good clinical application prospect.

The present invention discloses a pegRNA, a GRAND editing editing system and an application for specifically identifying the polled site of cattle without horns. The present invention belongs to the field of gene editing technology. Based on the lead editing, the present invention constructs a GRAND editing editing system and designs a specific pegRNA and a GE-pegRNA expression vector, which can target the polled site of cattle without horns, and realizes the replacement of the 6 bp sequence between 1706055-1706060 bp and the 208 bp sequence between 1705837-1706044 bp of the cattle genome, breaking through the bottleneck that the PE editing system can only insert ≤44bp fragments and has low insertion efficiency, realizing the insertion-deletion mutation of the 202 bp repetitive sequence, improving the insertion efficiency, and greatly enriching the application scenarios of the gene editing technology of long fragment sequence insertion without donor insertion.

Embodiments of the present disclosure relate to a method, apparatus, device, storage medium and program product for data query. The method comprises: determining, based on relevance to query information, a relevant data set from candidate data sets, the relevant data set comprising a plurality of relevant data, the relevance of the plurality of relevant data to the query information exceeding a threshold; adjusting, based on a preset sequence insertion operation, an arrangement order of the plurality of relevant data in the relevant data set; determining, based on the adjusted arrangement order of the plurality of relevant data, a prompt word having a preset format, the prompt word having the preset format being at least associated with an insertion mode of the preset sequence insertion operation; and obtaining a query result for the query information by inputting the prompt word and the relevant data associated with the prompt word into a preset query model. Thus, the accuracy of the search result can be improved.

The invention discloses a linear RNA (Ribonucleic Acid) precursor, a circular RNA as well as a preparation method and application of the linear RNA precursor and the circular RNA. The linear RNA precursor comprises a 5'outer homologous arm, a 3 'intron, a 3' exon, a 5 'inner homologous arm, a 5' spacer, an IRES sequence, an insertion exogenous sequence GOI, a 3 'spacer, a 3' inner homologous arm, a 5 'exon, a 5' intron and a 3 'outer homologous arm from the 5'end to the 3' end. The DNA sequences of the 5'and 3 'outer homologous arms, the 5' and 3 'spacers and the 5' and 3 'inner homologous arms are sequences shown as Seq ID No.1 to Seq ID No.6 in sequence. Through optimization of the homologous arms on the inner side and the outer side, the splicing activity of the intron can be better exerted, and the cyclization efficiency is improved; through spacer optimization, translation efficiency, cyclization efficiency and stability are improved. According to the preparation method disclosed by the invention, through optimization of multiple steps, the cyclization efficiency and the stability, purity and quality of the circular RNA are improved, and large-scale production is easy to realize.

The present invention provides, in particular, a method for recombining a re-editable target locus within a cell, comprising inserting an exogenous re-editable template sequence into the target locus; a method for recombining a re-editable cell, comprising a re-editable template sequence having two adjacent homology arms, wherein the re-editable template sequence is inserted into the target locus by homologous recombination; and methods for using the same. In some embodiments, a re-editable template sequence is provided, which has virtually no sequence similarity to any region in the genome, and which is recognizable by a genome editing system.

The present disclosure provides compositions and methods useful for inserting an exogenous donor DNA fragment into a target genomic sequence in a cell. The method comprises introducing two extension arms on a target genomic sequence and presenting to the genomic sequence a donor DNA having two complementary fragments flanking the donor fragment. The pairing of the extension arms allows donor DNA to be integrated into the genomic sequence. Depending on the configuration of the extension arm and a complementary fragment on the donor DNA, a portion of the genomic sequence may be repeated or deleted during this process.

The embodiment of the invention provides a microinstruction operation method and a processor. The method comprises the steps that the length of a to-be-inserted microinstruction sequence is determined; determining a target cache group according to the cache length of each cache group in the microinstruction cache; wherein the cache length of the target cache group is greater than or equal to the length of the microinstruction sequence; under the condition that the idle cache line exists in the target cache group, inserting the microinstruction sequence into the idle cache line; and under the condition that the idle cache line does not exist in the target cache group, determining a target cache line from the target cache group according to a target algorithm, and replacing the storage content in the target cache line with the microinstruction sequence. The cache length corresponding to each cache group in the microinstruction cache is set to be different, and the proper cache group is selected according to the sequence length of the microinstruction sequence, so that the waste of storage space is avoided, and the utilization rate of the microinstruction cache is improved.

The invention belongs to the field of gene engineering, and discloses an efficient and accurate plant gene knockout method. The method comprises the following steps: firstly, inserting a sequence containing a termination codon cluster into a genome through double pegRNA, so as to screen a termination codon cluster sequence capable of efficiently and accurately knocking out a target gene; in combination with the method, a guide editing system and a plurality of pegRNA programs are utilized, and various genome modifications such as efficient and accurate knockout, base replacement, small fragment insertion, deletion and replacement of one or more target genes can be realized by only one guide editing protein.