7 results about "Insertion deletion" patented technology

Provided is a reverse prime editing system, which is used for achieving site-specific and precise small-fragment DNA editing upstream of a target site. In the reverse prime editing system, a targeted strand nickase (such as D10A-Cas9) is used to cleave a nick on a targeted strand, thereby enabling precise small-fragment DNA base insertion, deletion and substitution upstream of the target site under the action of a reverse transcriptase.

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 provides an RNA framework for gene editing and a gene editing method, the RNA framework comprising a sequence upstream of a target site, a sequence to be inserted, and a sequence downstream of the target site in the 5' to 3' direction. The gene editing method is based on the inherent mechanism of eukaryotes, uses RNP or RNA (which can be prepared in vitro) and related proteins as carriers, and is transferred into the cytoplasm and nucleus to realize gene editing on a specific sequence or site on the genome of a target system, such as insertion, deletion, replacement and site replacement of a specific sequence, and has high targeting accuracy. The application is more suitable for further clinical application than other prior art because no exogenous system or substance such as protein derived from prokaryotes is introduced, and no double-strand break is generated.

This description relates to vectors for the genetic editing of filamentous fungi using the DNA double-strand break repair technique, capable of facilitating repair via homologous recombination, whether through the insertion, deletion or replacement of a DNA segment in the genome. These vectors preferably comprise the sequences defined in SEQ ID NO: 1 and SEQ ID NO: 23.

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.

The invention relates to a next-generation sequencing noise reduction method for simultaneously detecting BCR-ABL1 fusion and ABL1 mutation based on RNA and application, and belongs to the technical field of molecular diagnosis. The noise reduction method comprises the following steps: S1, carrying out reverse transcription on total RNA (Ribonucleic Acid) extracted from a leukemia sample to be detected to obtain a cDNA (Complementary Deoxyribonucleic Acid) product; s2, carrying out multiple PCR (Polymerase Chain Reaction) amplification to obtain DNA (Deoxyribonucleic Acid) subjected to PCR amplification; s3, performing linker connection, library construction and next-generation sequencing to obtain DNA library total quantity and sequencing data; and S4, carrying out raw signal filtering analysis, filtering false positive sites in a non-target region and background signals caused by reverse transcription, and synchronously judging a BCR-ABL1 fusion type, an ABL1 kinase region and point mutation and insertion deletion of a BCR gene. The noise reduction method can effectively reduce the interference of the sequencing background, not only expands the detection coverage range, but also has the technical advantages of simplicity and convenience in operation, excellent detection performance, low cost and high throughput.

The invention belongs to the field of gene engineering. The invention relates to a reverse guidance editing system, and particularly, the reverse guidance editing system is used for realizing fixed-point and accurate small-fragment DNA editing at the upstream of a target site. More specifically, according to the reverse guide editing system disclosed by the invention, a nick is cut on a targeting chain by utilizing targeting chain nicking enzyme (such as D10A-Cas9), and accurate small-fragment DNA base insertion, deletion and replacement are performed on the upstream of a target site under the action of reverse transcriptase; as well as genetically modified but non-transgenic organisms and progeny thereof produced by the method.