A list of the latest advances in gene editing technology in 2018.

The genome editing technology CRISPR/Cas9 has maintained rapid development since it was named one of science's top ten technology advances in 2013, with more than 51,700 crispR/Cas9-related documents on Google Academics.
to focus on the international and domestic cutting-edge gene editing technology, March 30-31, by Peking University National Key Laboratory on Natural Drugs and BionicDrugs, the 2018 Gene Editing Symposium will be held in Beijing.
will be a large number of first-line scientific research workers will gather here for a large academic feast.
over the past year, gene editing has continued to grow at a high rate, such as the discovery of proteins that can terminate the gene editing process, the manipulation of Cas9's REC3 domain to significantly reduce the off-target effect of CRISPR-Cas9, crispR gene editing combined with offliorgan organ construction techniques to help detect genetic cancer-specific DNA defects, and CRISPR's equipped phage to allow "superbugs" to commit suicide.
Wang Weiyi The application of highly effective and accurate genetic engineering techniques in animal model construction, T-cell therapy and gene expression regulation of the Institute of Zoology of the Chinese Academy of Sciences is of great value for the development of gene therapy and the establishment of cell and animal disease models.
has made great progress in the study of specific fixed-point nucleases in recent years.
in order to break through the limitations of traditional gene targeting methods, we were the first to obtain multi-gene knockout mice through the injection of CRISPR-Cas9 in fertilized eggs.
to replace low-throughput and highly technically demanding microinjection technology, we have established a method of electroshock of fertilized eggs, which greatly simplifies the construction of animal models.
this series of work has greatly improved the rate and speed of the building efficiency of genetically modified mice, and provided an important tool for invivia research in disease and developmental biology.
at the same time, we have established efficient single-gene and multigene knockout techniques in primary T cells and T-cells (CARTs) that express nhimtonoanti-disputing receptors, providing a tool for cell immunotherapy research.
In addition to gene editing, we have also established new technologies based on the CRISPR-Cas9 system to regulate gene expression levels and epigenetic modification.
Wang Yanli, The Institute of Biophysics of the Chinese Academy of Sciences, Cas13a, is a molecular mechanism for cutting RNA, the CRISPR/Cas system, a pronuclear bioimmune defense system that is the immune system mediated by small molecule RNA, which has been discovered in recent years.
CRISPR-Cas system is divided into two categories, Cas13a is the second class of VI-type system effect protein, also known as C2c2, with RNA-mediated RNA enzyme cutting activity, is currently the second class of CRISPR-Cas system found the only protein that can degrade RNA, in RNA technology has potential application value, the development of RNA tools, expanded CRISPR system in gene editing is of great value.
To study how Cas13a is activated to cut RNA, we analyzed the crystal structure of Leptotrichia Buccalis (Lbu) Cas13a, which binds to crRNA and its target RNA, and the cryo-EM structure of the Lbu Cas13a-crRNA complex.
results reveal edified that crRNA-target RNA bistrands bind to positively charged central channels of nuclease (NUC) blades, and that Cas13a proteins and crRNA seishaving significantly after target RNA binding.
guide the target RNA double-stranded formation to trigger the HEPN1 domain to move towards the HEPN2 domain, activating the HEPN catalytic site of the Cas13a protein, which is followed by nonspecific lysis of single-stranded targets and side-side RNAs.
results confirm that the binding of target RNA led to conformational changes in the two HEPN domains of LbuCas13a, which stimulated the enzyme cutting activity of LbuCas13a to cut any single-stranded RNA nonspecifically, and provided an important structural biological basis for the study of the molecular mechanisms of Cas13a's RNA enzyme activity.
the discovery of this study provides a reliable structural basis for the further development of the CRISPR-Cas13a system, provides strong evidence for a deep understanding of the molecular mechanism of bacteria to resist virus invasion, and will have great significance for the prevention, detection, control and treatment of virus-induced diseases, especially the effective RNA enzyme cutting activity based on Cas13a, which has a very broad prospect for its rapid detection of various major diseases.
Wu Qiang Shanghai Jiaotong University developed DNA large fragment editing technology to study the advanced structure of chromatin derived from bacteria and archaea's type II cluster regular interval short-text repetition system .Clustered regularly spaced palin shortdromic repeats (CRISPR)/CRISPR-associated nuclease 9 (Cas9), CRISPR/Cas9 has been transformed into a new technology for fixed-point genome editing in recent years.
because of its simple design, easy to operate, low cost and other great advantages, to the field of genetic operation has brought about a revolutionary change.
the targeted editing technique of genomic DNA fragments based on CRISPR/Cas9 system mainly includes the deletion, inversion, repetition, insertion and translocation of DNA fragments, and this effective DNA fragment editing method provides a powerful means for studying gene function, regulatory elements, tissue development and disease development.
I will highlight our latest research in this field on the evolution of chromatin architecture proteins such as CTCF in the regulation of 3D genome assembly, providing a reference for gene regulation and functional research using genomic DNA fragments for targeted editing.
Zhou Demin Beijing University National Key Laboratory for Natural Drugs and Bionic Drugs The conversion of life-bully viruses into live but avirulent vaccines a revolution in vaccinology. In a proof-principle study, we're swydd the sylwedd the code of the sinoa virus a transgenic cell line-in-the-orthogonal translation machinery. This generated dydd dydd dydd (PTC) - harboring viruses that exerted full infectivity byison wasa iad a tha a' sad a' saper-in-a-sa- saud. Genome-wide optimization of the sites for the incorporation of multiple PTCs resulted in highly ar y mondwl styroth sy'r sions in transgenic cells. In mouse, ferret, and pig models, vaccine with PTC viruses elic robustsaude, mucosal, and T cell- Wang Yongming Fudan University to establish an efficient CRISPR/Cas9 editing technology CRISPR/Cas9 is a revolutionary gene editing technology that is widely used.
we're working on two things to improve the editing efficiency of CRISPR/Cas9.
we use attachment vectors to express Cas9 and gRNA, known as epiCRISPR technology.
attachment vectors are not integrated into the genome, but can be replicated as cells divide, so they can express exogenous genes over a long period of time and achieve long-term gene editing.
at the same time on the attachment carrier to express the anti-mycin resistance gene, can enrich the successful transfection of cells.
remove the screening drug after the editing, attach the body granules are rapidly lost, and the edited cells no longer express the exogenous gene.
can achieve up to 100% editing efficiency with attachment technology, and efficiently knock out large fragments of the genome, as well as knock out multiple genes at the same time.
CRISPR/Cas9 editing efficiency is affected by gRNA sequences, the efficiency of different gRNA varies greatly, and testing gRNA efficiency is time-consuming and laborious.
we established a high-flow method to test gRNA activity, and obtained more than 50,000 gRNA activity, covering more than 20,000 human genes.
these efforts will greatly facilitate gene editing.
Changxing Shanghai Institute of Life Sciences, Chinese Academy of Sciences, uses targeted cytosine deaminase for gene editing of mononucleotided diversity is the main source of genetic diversity, an important genetic basis for human individual differences, but also the driving force of molecular evolution and a direct cause of many diseases.
, however, in mammals, there is still a lack of tools to effectively induce mutations of single nucleotides, and the function of studying single nucleotide mutations in experimental efficiency and high-volume.
most of the existing experimental techniques can only disrupt the function or expression of genes, resulting in the lack of gene function, to induce the acquisition of new functions can not be done.
using targeted cytosine deaminase-mediated base editing (Targeted-mediated mutagenesis, TAM) technology, the cytosine and ostrich can be randomly transformed into the other three bases on the targeted genomic DNA of sgRNA, resulting in massive mutants, combined with screening, to analyze the function of single nucleotide mutations or induce the evolution of proteins in the body.
at the same time with the assistance of a polypeptide inhibitor (UGI), dCas9-AID can induce a specific cytosine transition to thymus, enable precise editing of a single base, and provide a solution for the treatment of genetic diseaseinduced by single nucleotide mutations.
using this technology, it has been successfully screened for reported and new imatinib drug resistance sites in chronic myeloma cells.
, as a new technique of efficient mammalian DNA base editing, TAM can be widely used in protein engineering, molecular genetics research and gene therapy.
Zhu Jie Guangzhou Women's and Children's Medical Center CRISPR-Cas9-mediated photoreceptor cell reprogramming therapy of retinal pigmentation gene therapy has shown great potential in the treatment of many human diseases.
However, the current method is usually aimed at only a single gene or a single mutation, which greatly limits the treatment of multi-gene or multi-point mutations.
retinal pigmentation is a common clinical blinding eye disease, which is very clinical and genetic heterogeneity, and is one of the most important diseases leading to human vision impairment.
more than 40 genes and sites have been reported to be associated with retinal pigmentation.
here we used CRISPR/Cas9-mediated gene editing methods to treat retinal pigmentation mice.
reprogramthetic cell reprogramming as a cone cell by mutationther the important transcription factor in the rod cell, NRL or Nr2e3.
converted cone-like cells are insensitive to the degradation of photosensitive cells caused by mutations, so that mice can retain a certain degree of vision later in the disease.
our findings not only provide new, non-gene-dependent treatments for retinal pigmentation, but also demonstrate the great possibilities for gene reprogramming, cell degradation prevention, and tissue function protection mediated by gene editing techniques.
Yang Hui , the application of gene modification animals in disease modeling and disease treatment by gene editing of the Shanghai Institute of Neurology of the Chinese Academy of Sciences is an important tool for studying gene function in development and disease .
CRISPR/Cas9 system is effectively used to build gene knockout and knock-in mice.
however, the genetically modified animals obtained by this method have a serious chimeric phenomenon, in which one part of the individual animal is genetically edited and the other part does not. it takes a long time and a lot of time to knock out mice
the gene that obtains the purification through mating, especially in the mice that obtainmultigene knockout.
because the monkeys had a long reproductive cycle (4-5 years of sexual maturity, half a year of pregnancy), low reproductive capacity (monoletarys), it took longer and cost to obtain genetically modified monkeys with pure mutations through mating methods.
For this reason, by optimizing the CRISPR/Cas9 system, we have successfully obtained single-gene or multigene function in the first generation of mice and monkeys, which can be used directly for phenotype analysis, greatly promoting the establishment of non-human primate models and their research in brain science and brain diseases.
we have also designed a homologous-mediated end joint (HMEJ) strategy that enables precise and efficient gene integration in both divided and non-divided cells.
more important, the method was much more efficient than the HR, NHEJ, and MMEJ-based strategies in mouse and monkey embryos or liver cells and neurons in the body.
, the HMEJ strategy may be of many uses, such as gene editing to obtain animal models and targeted gene therapy.
through these methods, we can effectively obtain a variety of genetically modified monkey models in monkeys.
recently, our target sedation of disease monkey models includes PD, AD, ALS, DMD, RP, AS, etc., and tool monkey models include photogenetic monkeys, various neuron-specific Cre monkeys, and so on.