CRISPR Journal最新文献_第10页

Prime Editing in Mammals: The Next Generation of Precision Genome Editing. 哺乳动物的初始编辑:下一代精确基因组编辑。

IF 3.7 4区生物学

CRISPR Journal Pub Date : 2022-12-01 DOI: 10.1089/crispr.2022.0084

Dawei Wang, Xiude Fan, Mengzhu Li, Tianbo Liu, Peng Lu, Guangxin Wang, Yuan Li, JunMing Han, JiaJun Zhao

引用次数: 0

CRISPR-Mediated Cassette Exchange (CriMCE): A Method to Introduce and Isolate Precise Marker-Less Edits. crispr介导的盒式交换(crice):一种引入和分离精确的无标记编辑的方法。

IF 3.7 4区生物学

CRISPR Journal Pub Date : 2022-12-01 DOI: 10.1089/crispr.2022.0026

Ioanna Morianou, Andrea Crisanti, Tony Nolan, Andrew M Hammond

引用次数: 1

Impact of KIT Editing on Coat Pigmentation and Fresh Meat Color in Yorkshire Pigs. KIT编辑对约克郡猪被毛色素沉着和鲜肉颜色的影响。

IF 3.7 4区生物学

CRISPR Journal Pub Date : 2022-12-01 DOI: 10.1089/crispr.2022.0039

Xinyu Liang, Jin Lan, Meina Xu, Ke Qin, Hongbo Liu, Guanjie Sun, Xiaohong Liu, Yaosheng Chen, Zuyong He

{"title":"Impact of KIT Editing on Coat Pigmentation and Fresh Meat Color in Yorkshire Pigs.","authors":"Xinyu Liang, Jin Lan, Meina Xu, Ke Qin, Hongbo Liu, Guanjie Sun, Xiaohong Liu, Yaosheng Chen, Zuyong He","doi":"10.1089/crispr.2022.0039","DOIUrl":"https://doi.org/10.1089/crispr.2022.0039","url":null,"abstract":"The white coat color of Yorkshire pigs is caused by the dominant white I allele, which has been associated with at least one copy of the 450-kb duplication encompassing the entire KIT gene and a splice mutation (G > A) at the first base of intron 17. The splice mutation in KIT has an adverse effect on pigmentation in mice. Therefore, removing the 450 kb duplications harboring the KIT copy with splice mutations is expected to affect Yorkshire pig pigmentation. In this study, we describe the use of a Yorkshire pig kidney cell strain with the I?/IBe-ed genotype, previously created by CRISPR-Cas9, as donor cells for somatic cell nuclear transfer to generate gene-edited Yorkshire pigs. The removal of the 450 kb duplications harboring the KIT copy with splice mutation did not alter the white coat color of Yorkshire pigs, which was confirmed by the absence of fully mature melanocytes and melanin accumulation in the hair follicles. Except for the improved transcription of tyrosinase, and slight increase in microphthalmia transcription factor and tyrosinase-related protein 1 protein expression, there was no significant impact of the removal of splice mutations on genes and signaling pathways (PI3K/AKT) involved in melanogenesis. However, the removal of the 450 kb duplications harboring the KIT copy with splice mutation substantially improved fresh meat color accompanied by significantly increased red blood cell number, which merits further investigation. Our study provides new insights into the role of structural mutations of the KIT gene in the formation of white coat color and erythropoiesis in Yorkshire pigs.","PeriodicalId":54232,"journal":{"name":"CRISPR Journal","volume":null,"pages":null},"PeriodicalIF":3.7,"publicationDate":"2022-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"9187188","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Natural Nucleoside Modifications in Guide RNAs Can Modulate the Activity of the CRISPR-Cas9 System In Vitro. 引导rna中的天然核苷修饰可以在体外调节CRISPR-Cas9系统的活性

IF 3.7 4区生物学

CRISPR Journal Pub Date : 2022-12-01 DOI: 10.1089/crispr.2022.0069

Daria V Prokhorova, Ivan P Vokhtantsev, Polina O Tolstova, Evgenii S Zhuravlev, Lilia M Kulishova, Dmitry O Zharkov, Grigory A Stepanov

{"title":"Natural Nucleoside Modifications in Guide RNAs Can Modulate the Activity of the CRISPR-Cas9 System In Vitro.","authors":"Daria V Prokhorova, Ivan P Vokhtantsev, Polina O Tolstova, Evgenii S Zhuravlev, Lilia M Kulishova, Dmitry O Zharkov, Grigory A Stepanov","doi":"10.1089/crispr.2022.0069","DOIUrl":"https://doi.org/10.1089/crispr.2022.0069","url":null,"abstract":"At the present time, the clustered regularly interspaced short palindromic repeats (CRISPR)-CRISPR-associated protein 9 (Cas9) system has been widely adopted as an efficient genomic editing tool. However, there are some actual problems such as the off-target effects, cytotoxicity, and immunogenicity. The incorporation of modifications into guide RNAs permits enhancing both the efficiency and the specificity of the CRISPR-Cas9 system. In this study, we demonstrate that the inclusion of N6-methyladenosine, 5-methylcytidine, and pseudouridine in trans-activating RNA (tracrRNA) or in single guide RNA (sgRNA) enables efficient gene editing in vitro. We found that the complexes of modified guide RNAs with Cas9 protein promoted cleavage of the target short/long duplexes and plasmid substrates. In addition, the modified monomers in guide RNAs allow increasing the specificity of CRISPR-Cas9 system in vitro and promote diminishing both the immunostimulating and the cytotoxic effects of sgRNAs.","PeriodicalId":54232,"journal":{"name":"CRISPR Journal","volume":null,"pages":null},"PeriodicalIF":3.7,"publicationDate":"2022-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"10079957","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 4

Bio-Orthogonal Chemistry Conjugation Strategy Facilitates Investigation of N-methyladenosine and Thiouridine Guide RNA Modifications on CRISPR Activity. 生物正交化学偶联策略促进了n -甲基腺苷和硫嘧啶引导RNA修饰对CRISPR活性的研究。

IF 3.7 4区生物学

CRISPR Journal Pub Date : 2022-12-01 DOI: 10.1089/crispr.2022.0065

Alyssa Hoy, Ya Ying Zheng, Jia Sheng, Maksim Royzen

{"title":"Bio-Orthogonal Chemistry Conjugation Strategy Facilitates Investigation of N-methyladenosine and Thiouridine Guide RNA Modifications on CRISPR Activity.","authors":"Alyssa Hoy, Ya Ying Zheng, Jia Sheng, Maksim Royzen","doi":"10.1089/crispr.2022.0065","DOIUrl":"https://doi.org/10.1089/crispr.2022.0065","url":null,"abstract":"The CRISPR-Cas9 system is an important genome editing tool that holds enormous potential toward the treatment of human genetic diseases. Clinical success of CRISPR technology is dependent on the incorporation of modifications into the single-guide RNA (sgRNA). However, chemical synthesis of modified sgRNAs, which are over 100 nucleotides in length, is difficult and low-yielding. We developed a conjugation strategy that utilized bio-orthogonal chemistry to efficiently assemble functional sgRNAs containing nucleobase modifications. The described approach entails the chemical synthesis of two shorter RNA oligonucleotides: a 31-mer containing tetrazine (Tz) group and a 70-mer modified with a trans-cyclooctene (TCO) moiety. The two oligonucleotides were conjugated to form functional sgRNAs. The two-component conjugation methodology was utilized to synthesize a library of sgRNAs containing nucleobase modifications such as N1-methyladenosine (m1A), N6-methyladenosine (m6A), 2-thiouridine (s2U), and 4-thiouridine (s4U). The impact of these RNA modifications on overall CRISPR activity were investigated in vitro and in Cas9-expressing HEK293T cells.","PeriodicalId":54232,"journal":{"name":"CRISPR Journal","volume":null,"pages":null},"PeriodicalIF":3.7,"publicationDate":"2022-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9805849/pdf/crispr.2022.0065.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"10079973","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 2

Highly Efficient One-Step Tagging of Endogenous Genes in Primary Cells Using CRISPR-Cas Ribonucleoproteins. 利用CRISPR-Cas核糖核蛋白高效一步标记原代细胞内源基因。

IF 3.7 4区生物学

CRISPR Journal Pub Date : 2022-12-01 DOI: 10.1089/crispr.2022.0046

Yao Yao, Jiaxuan Cao, Wentian Wang, Boya Liu, Xiaolei Pei, Lei Zhang, Shuquan Rao

引用次数: 0

Better Guidance for CRISPR. 更好地指导CRISPR。

IF 3.7 4区生物学

CRISPR Journal Pub Date : 2022-12-01 DOI: 10.1089/crispr.2022.29156.editorial

Rodolphe Barrangou

引用次数: 0

Increasing Genome Editing Efficiency of Cas9 Nucleases by the Simultaneous Use of Transcriptional Activators and Histone Acetyltransferase Activator. 同时使用转录激活剂和组蛋白乙酰转移酶激活剂提高Cas9核酸酶的基因组编辑效率

IF 3.7 4区生物学

CRISPR Journal Pub Date : 2022-12-01 DOI: 10.1089/crispr.2022.0001

Junhao Liu, Bo Li, Lele Yang, Naixia Ren, Meichen Xu, Qilai Huang

{"title":"Increasing Genome Editing Efficiency of Cas9 Nucleases by the Simultaneous Use of Transcriptional Activators and Histone Acetyltransferase Activator.","authors":"Junhao Liu, Bo Li, Lele Yang, Naixia Ren, Meichen Xu, Qilai Huang","doi":"10.1089/crispr.2022.0001","DOIUrl":"https://doi.org/10.1089/crispr.2022.0001","url":null,"abstract":"The CRISPR-Cas9 system shows diverse levels of genome editing activities on eukaryotic chromatin, and high-efficiency sgRNA targets are usually desired in application. In this study, we show that chromatin open status is a pivotal determinant of the Cas9 editing activity in mammalian cells, and increasing chromatin accessibility can efficiently improve Cas9 genome editing. However, the strategy that increases chromatin openness by fusing the VP64 transcriptional activation domain at the C-terminus of Cas9 can only promote genome editing activity slightly at most tested CRISPR-Cas9 targets in Lenti-X 293T cells. Under the enlightenment that histone acetylation increases eukaryotic chromatin accessibility, we developed a composite strategy to further improve genome editing by activating histone acetylation. We demonstrate that promoting histone acetylation using the histone acetyltransferase activator YF-2 can improve the genome editing by Cas9 and, more robustly, by the Cas9 transcriptional activator (Cas9-AD). This strategy holds great potential to enhance CRISPR-Cas9 genome editing and to enable broader CRISPR gRNA target choices for experiments in eukaryotes.","PeriodicalId":54232,"journal":{"name":"CRISPR Journal","volume":null,"pages":null},"PeriodicalIF":3.7,"publicationDate":"2022-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"10624510","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 2

A Novel CRISPR Interference Effector Enabling Functional Gene Characterization with Synthetic Guide RNAs. 一种新的CRISPR干扰效应物，可以用合成的引导rna来表征功能基因。

IF 3.7 4区生物学

CRISPR Journal Pub Date : 2022-12-01 Epub Date: 2022-10-17 DOI: 10.1089/crispr.2022.0056

Clarence Mills, Andrew Riching, Ashleigh Keller, Jesse Stombaugh, Amanda Haupt, Elena Maksimova, Sarah M Dickerson, Emily Anderson, Kevin Hemphill, Chris Ebmeier, John A Schiel, Josien Levenga, Matthew Perkett, Anja van Brabant Smith, Zaklina Strezoska

{"title":"A Novel CRISPR Interference Effector Enabling Functional Gene Characterization with Synthetic Guide RNAs.","authors":"Clarence Mills, Andrew Riching, Ashleigh Keller, Jesse Stombaugh, Amanda Haupt, Elena Maksimova, Sarah M Dickerson, Emily Anderson, Kevin Hemphill, Chris Ebmeier, John A Schiel, Josien Levenga, Matthew Perkett, Anja van Brabant Smith, Zaklina Strezoska","doi":"10.1089/crispr.2022.0056","DOIUrl":"10.1089/crispr.2022.0056","url":null,"abstract":"While CRISPR interference (CRISPRi) systems have been widely implemented in pooled lentiviral screening, there has been limited use with synthetic guide RNAs for the complex phenotypic readouts enabled by experiments in arrayed format. Here we describe a novel deactivated Cas9 fusion protein, dCas9-SALL1-SDS3, which produces greater target gene repression than first or second generation CRISPRi systems when used with chemically modified synthetic single guide RNAs (sgRNAs), while exhibiting high target specificity. We show that dCas9-SALL1-SDS3 interacts with key members of the histone deacetylase and Swi-independent three complexes, which are the endogenous functional effectors of SALL1 and SDS3. Synthetic sgRNAs can also be used with in vitro-transcribed dCas9-SALL1-SDS3 mRNA for short-term delivery into primary cells, including human induced pluripotent stem cells and primary T cells. Finally, we used dCas9-SALL1-SDS3 for functional gene characterization of DNA damage host factors, orthogonally to small interfering RNA, demonstrating the ability of the system to be used in arrayed-format screening.","PeriodicalId":54232,"journal":{"name":"CRISPR Journal","volume":null,"pages":null},"PeriodicalIF":3.7,"publicationDate":"2022-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9805873/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"9199643","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

CRISPR-Cas9 Genome Editing Uncovers the Mode of Action of Methoprene in the Yellow Fever Mosquito, Aedes aegypti. CRISPR-Cas9基因组编辑揭示了甲氧丁烯在黄热病蚊子，埃及伊蚊中的作用模式。

IF 3.7 4区生物学

CRISPR Journal Pub Date : 2022-12-01 DOI: 10.1089/crispr.2022.0066

Guan-Heng Zhu, Sharath Chandra Gaddelapati, Yaoyu Jiao, Jinmo Koo, Subba Reddy Palli

{"title":"CRISPR-Cas9 Genome Editing Uncovers the Mode of Action of Methoprene in the Yellow Fever Mosquito, Aedes aegypti.","authors":"Guan-Heng Zhu, Sharath Chandra Gaddelapati, Yaoyu Jiao, Jinmo Koo, Subba Reddy Palli","doi":"10.1089/crispr.2022.0066","DOIUrl":"https://doi.org/10.1089/crispr.2022.0066","url":null,"abstract":"Methoprene, a juvenile hormone (JH) analog, is widely used for insect control, but its mode of action is not known. To study methoprene action in the yellow fever mosquito, Aedes aegypti, the E93 (ecdysone-induced transcription factor) was knocked out using the CRISPR-Cas9 system. The E93 mutant pupae retained larval tissues similar to methoprene-treated insects. These insects completed pupal ecdysis and died as pupa. In addition, the expression of transcription factors, broad complex and Krüppel homolog 1 (Kr-h1), increased and that of programmed cell death (PCD) and autophagy genes decreased in E93 mutants. These data suggest that methoprene functions through JH receptor, methoprene-tolerant, and induces the expression of Kr-h1, which suppresses the expression of E93, resulting in a block in PCD and autophagy of larval tissues. Failure in the elimination of larval tissues and the formation of adult structures results in their death. These results answered long-standing questions on the mode of action of methoprene.","PeriodicalId":54232,"journal":{"name":"CRISPR Journal","volume":null,"pages":null},"PeriodicalIF":3.7,"publicationDate":"2022-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9805843/pdf/crispr.2022.0066.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"9200163","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0