Experimental
Teaching Design for Gene Editing in Rice Breeding by CRISPR/Cas9 Technology

doi:10.11983/CBB25080

Abstract

Abstract: CRISPR/Cas9 technology, as an efficient genome editing tool, has gained increasingly widespread application in the life sciences. Incorporating this technology into university experimental teaching enables students to deepen their understanding of cutting-edge research advances while enhancing their innovative research capabilities and practical skills. In this paper, we have designed an innovative teaching experiment, “Using CRISPR/Cas9 technology to study the effect of knocking out rice SSA1(seedling stage albino1)gene on phenotype”. This experiment helps students to deepen their understanding of the principles and experimental procedures of CRISPR/Cas9 technology and to recognize the important role played by theSSA1 gene in rice growth and development. At the same time, the experiment can improve students' ability of experimental operation and rigorous scientific research thinking, and cultivate students' exploratory skills and innovative spirit.

Key words: CRISPR/Cas9,, gene editing,, rice,, experimental instructional design

Ruotong Lü, Yunxia Kou, Yujia Xiao, Pengfei Ai, Yanwei Wang.

Experimental Teaching Design for Gene Editing in Rice Breeding by CRISPR/Cas9 Technology[J]. Chinese Bulletin of Botany, DOI: 10.11983/CBB25080.

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URL: https://www.chinbullbotany.com/EN/10.11983/CBB25080

References

[1]Jansen R, Embden JD, Gaastra W, Schouls LM(2002).Identification of genes that are associated with DNA repeats in prokaryotes.Molecular microbiology, 43:1565-1575. [2]Ishino Y, Shinagawa H, Makino K, Amemura M, Nakata A(1987).Nucleotide sequence of the iap gene,responsible for alkaline phosphatase isozyme conversion in Escherichia coli,and identification of the gene product.Journal of bacteriology, 169:5429-5433. [3]Mojica FJ, Díez-Villase?or C, Soria E, Juez G(2000).Biological significance of a family of regularly spaced repeats in the genomes of Archaea,Bacteria and mitochondria.Molecular microbiology, 36:244-246. [4]Bolotin A, Quinquis B, Sorokin A, Ehrlich SD(2005).Clustered regularly interspaced short palindrome repeats (CRISPRs) have spacers of extrachromosomal origin.Microbiology (Reading, England) 151(Pt, 151:2551-2561. [5]Horvath P, Barrangou R(2010).CRISPRCas,the immune system of bacteria and archaea.Science (New York, N Y), 327:167-170. [6]Semenova E, Jore MM, Datsenko K A, Semenova A, Westra ER, Wanner B, van der Oost J, Brouns SJ, Severinov K(2011).Interference by clustered regularly interspaced short palindromic repeat (CRISPR) RNA is governed by a seed sequence.PNAS, 108:10098-10103. [7]Koonin EV, Wolf YI(2009).Is evolution Darwinian or/and Lamarckian?.Biology direct, :42-42. [8]Sander JD, Dahlborg EJ, Goodwin MJ, Cade L, Zhang F, Cifuentes D, Curtin SJ, Blackburn JS, Thibodeau-Beganny S, Qi Y, Pierick CJ, Hoffman E, Maeder M L, Khayter C, Reyon D, Dobbs D, Langenau DM, Stupar RM, Giraldez AJ, Voytas DF, Joung JK(2011).Selection-free zinc-finger-nuclease engineering by context-dependent assembly (CoDA).Nature methods, 8:67-69. [9]Reyon D, Tsai S Q, Khayter C, Foden JA, Sander JD, Joung JK(2012).FLASH assembly of TALENs for high-throughput genome editing.Nature biotechnology, 30:460-465. [10]McManus MT, Sharp PA(2002).Gene silencing in mammals by small interfering RNAs.Nature reviews. Genetics, 3:737-747. [11]Hsu P D, Scott D A, Weinstein J A, Ran F A, Konermann S, Agarwala V, Li Y, Fine EJ, Wu X, Shalem O, Cradick TJ, Marraffini LA, Bao G, Zhang F(2013).DNA targeting specificity of RNA-guided Cas9 nucleases.Nature biotechnology, 31:827-832. [12]Pattanayak V, Lin S, Guilinger JP, Ma E, Doudna JA, Liu DR(2013).High-throughput profiling of off-target DNA cleavage reveals RNA-programmed Cas9 nuclease specificity.Nature biotechnology, 31:839-843. [13]IzawaT, Shimamoto K(1996).Becoming a model plant: The importance of rice to plant science.Trends Plant Sci, 1:95-99. [14]Gale MD, Devos KM(1998).Comparative genetics in the grasses.PNAS, 95:1971-1974. [15]Moore G, Devos KM, Wang Z, Gale MD(1995).Cereal genome evolution.Grasses, line up and form a circle. Current biology : CB, 5:737-739. [16]Wang Y, Yang Z, Zhang M, Ai P(2022).A chloroplast-localized pentatricopeptide repeat protein involved in RNA editing and splicing and its effects on chloroplast development in rice..BMC plant biology, 22:437-437. [17]Jinek M, Chylinski K, Fonfara I, Hauer M, Doudna JA, Charpentier E(2012).A programmable dual-RNA-guided DNA endonuclease in adaptive bacterial immunity.Science (New York, N.Y.), 337:816-821. [18]Jinek M, Jiang F, Taylor DW, Sternberg SH, Kaya E, Ma E, Anders C, Hauer M, Zhou K, Lin S, Kaplan M, Iavarone AT, Charpentier E, Nogales E, Doudna JA(2014).Structures of Cas9 endonucleases reveal RNA-mediated conformational activation..Science (New York, N.Y.), 343:1247997-. [19]Nishimasu H, Ran FA, Hsu PD, Konermann S, Shehata SI, Dohmae N, Ishitani R, Zhang F, Nureki O(2014).Crystal structure of Cas9 in complex with guide RNA and target DNA.Cell, 156:935-949. [20]Wyman C, Kanaar R(2006).DNA double-strand break repair: all's well that ends well.Annual review of genetics, 40:363-383. [21]Baltes NJ, Voytas DF(2015).Enabling plant synthetic biology through genome engineering.Trends in biotechnology, 33:120-131. [22]Wang J, Palomar VM, Min JH, Wierzbicki AT(2024).PAP1 and PAP7 are required for association of plastid-encoded RNA polymerase with DNA..Plant molecular biology, 114:100-100. [23]曾栋昌, 马兴亮, 谢先荣, 祝钦泷, 刘耀光(2018).植物多基因编辑载体构建和突变分析的操作方法.中国科学生命科学, 48:783-794. [24]Ma X, Zhang Q, Zhu Q, Liu W, Chen Y, Qiu R, Wang B, Yang Z, Li H, Lin Y, Xie Y, Shen R, Chen S, Wang Z, Chen Y, Guo J, Chen L, Zhao X, Dong Z, Liu YG(2015).A Robust CRISPRCas9 System for Convenient,High-Efficiency Multiplex Genome Editing in Monocot and Dicot Plants.Molecular plant, 8:1274-1284. [25]高凌飚(2004).关于过程性评价的思考.课程.教材.教法, 10:15-19.

Experimental Teaching Design for Gene Editing in Rice Breeding by CRISPR/Cas9 Technology

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