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Engineering T Cells Using CRISPR/Cas9 for Cancer Therapy

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RNA Interference and CRISPR Technologies

Part of the book series: Methods in Molecular Biology ((MIMB,volume 2115))

Abstract

Recent advances in the development of gene editing technologies, especially the CRISPR/Cas 9 system, have substantially enhanced our ability to make precise and efficient changes in the genomes of various cells. In particular, the genetic engineering of T cells holds huge potential to improve the efficacy and safety of T cells-based cancer therapy. Due to its ease of use and high efficiency, CRISPR/Cas9 enables efficient gene knockout, site-specific knock-in, and genome-wide screen in T cells. Here we review the current progress of applying gene editing to T-cell therapy, focusing on the technical aspects of the CRISPR/Cas9 platform. We also discuss the challenges and future prospects.

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References

  1. Rouet P, Smih F, Jasin M (1994) Introduction of double-strand breaks into the genome of mouse cells by expression of a rare-cutting endonuclease. Mol Cell Biol 14:8096–8106

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  2. Miller JC, Holmes MC, Wang J, Guschin DY, Lee YL, Rupniewski I, Beausejour CM, Waite AJ, Wang NS, Kim KA, Gregory PD, Pabo CO, Rebar EJ (2007) An improved zinc-finger nuclease architecture for highly specific genome editing. Nat Biotechnol 25:778–785

    Article  CAS  PubMed  Google Scholar 

  3. Wolfe SA, Nekludova L, Pabo CO (2000) DNA recognition by Cys2His2 zinc finger proteins. Annu Rev Biophys Biomol Struct 29:183–212

    Article  CAS  PubMed  Google Scholar 

  4. Urnov FD, Miller JC, Lee YL, Beausejour CM, Rock JM, Augustus S, Jamieson AC, Porteus MH, Gregory PD, Holmes MC (2005) Highly efficient endogenous human gene correction using designed zinc-finger nucleases. Nature 435:646–651

    Article  CAS  PubMed  Google Scholar 

  5. Porteus MH, Baltimore D (2003) Chimeric nucleases stimulate gene targeting in human cells. Science 300:763

    Article  PubMed  Google Scholar 

  6. Miller J, McLachlan AD, Klug A (1985) Repetitive zinc-binding domains in the protein transcription factor IIIA from Xenopus oocytes. EMBO J 4:1609–1614

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  7. Bibikova M, Golic M, Golic KG, Carroll D (2002) Targeted chromosomal cleavage and mutagenesis in drosophila using zinc-finger nucleases. Genetics 161:1169–1175

    CAS  PubMed  PubMed Central  Google Scholar 

  8. Bibikova M, Beumer K, Trautman JK, Carroll D (2003) Enhancing gene targeting with designed zinc finger nucleases. Science 300:764

    Article  CAS  PubMed  Google Scholar 

  9. Miller JC, Tan S, Qiao G, Barlow KA, Wang J, Xia DF, Meng X, Paschon DE, Leung E, Hinkley SJ, Dulay GP, Hua KL, Ankoudinova I, Cost GJ, Urnov FD, Zhang HS, Holmes MC, Zhang L, Gregory PD, Rebar EJ (2011) A TALE nuclease architecture for efficient genome editing. Nat Biotechnol 29:143–148

    Article  CAS  PubMed  Google Scholar 

  10. Li T, Huang S, Jiang WZ, Wright D, Spalding MH, Weeks DP, Yang B (2011) TAL nucleases (TALNs): hybrid proteins composed of TAL effectors and FokI DNA-cleavage domain. Nucleic Acids Res 39:359–372

    Article  PubMed  CAS  Google Scholar 

  11. Christian M, Cermak T, Doyle EL, Schmidt C, Zhang F, Hummel A, Bogdanove AJ, Voytas DF (2010) Targeting DNA double-strand breaks with TAL effector nucleases. Genetics 186:757–761

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  12. Boch J, Scholze H, Schornack S, Landgraf A, Hahn S, Kay S, Lahaye T, Nickstadt A, Bonas U (2009) Breaking the code of DNA binding specificity of TAL-type III effectors. Science 326:1509–1512

    Article  CAS  PubMed  Google Scholar 

  13. Mali P, Yang L, Esvelt KM, Aach J, Guell M, DiCarlo JE, Norville JE, Church GM (2013) RNA-guided human genome engineering via Cas9. Science 339:823–826

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  14. Cong L, Ran FA, Cox D, Lin S, Barretto R, Habib N, Hsu PD, Wu X, Jiang W, Marraffini LA, Zhang F (2013) Multiplex genome engineering using CRISPR/Cas systems. Science 339:819–823

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  15. Gasiunas G, Barrangou R, Horvath P, Siksnys V (2012) Cas9-crRNA ribonucleoprotein complex mediates specific DNA cleavage for adaptive immunity in bacteria. Proc Natl Acad Sci U S A 109:E2579–E2586

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  16. 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 337:816–821

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  17. Sapranauskas R, Gasiunas G, Fremaux C, Barrangou R, Horvath P, Siksnys V (2011) The Streptococcus thermophilus CRISPR/Cas system provides immunity in Escherichia coli. Nucleic Acids Res 39:9275–9282

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  18. Deltcheva E, Chylinski K, Sharma CM, Gonzales K, Chao Y, Pirzada ZA, Eckert MR, Vogel J, Charpentier E (2011) CRISPR RNA maturation by trans-encoded small RNA and host factor RNase III. Nature 471:602–607

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  19. Garneau JE, Dupuis ME, Villion M, Romero DA, Barrangou R, Boyaval P, Fremaux C, Horvath P, Magadan AH, Moineau S (2010) The CRISPR/Cas bacterial immune system cleaves bacteriophage and plasmid DNA. Nature 468:67–71

    Article  CAS  PubMed  Google Scholar 

  20. Barrangou R, Fremaux C, Deveau H, Richards M, Boyaval P, Moineau S, Romero DA, Horvath P (2007) CRISPR provides acquired resistance against viruses in prokaryotes. Science 315:1709–1712

    Article  CAS  PubMed  Google Scholar 

  21. Bolotin A, Quinquis B, Sorokin A, Ehrlich SD (2005) Clustered regularly interspaced short palindrome repeats (CRISPRs) have spacers of extrachromosomal origin. Microbiology 151:2551–2561

    Article  CAS  PubMed  Google Scholar 

  22. Morgan RA, Dudley ME, Wunderlich JR, Hughes MS, Yang JC, Sherry RM, Royal RE, Topalian SL, Kammula US, Restifo NP, Zheng Z, Nahvi A, de Vries CR, Rogers-Freezer LJ, Mavroukakis SA, Rosenberg SA (2006) Cancer regression in patients after transfer of genetically engineered lymphocytes. Science 314:126–129

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  23. Wang M, Yin B, Wang HY, Wang RF (2014) Current advances in T-cell-based cancer immunotherapy. Immunotherapy 6:1265–1278

    Article  CAS  PubMed  Google Scholar 

  24. Turtle CJ, Hanafi LA, Berger C, Gooley TA, Cherian S, Hudecek M, Sommermeyer D, Melville K, Pender B, Budiarto TM, Robinson E, Steevens NN, Chaney C, Soma L, Chen X, Yeung C, Wood B, Li D, Cao J, Heimfeld S, Jensen MC, Riddell SR, Maloney DG (2016) CD19 CAR-T cells of defined CD4+:CD8+ composition in adult B cell ALL patients. J Clin Invest 126:2123–2138

    Article  PubMed  PubMed Central  Google Scholar 

  25. Maude SL, Laetsch TW, Buechner J, Rives S, Boyer M, Bittencourt H, Bader P, Verneris MR, Stefanski HE, Myers GD, Qayed M, De Moerloose B, Hiramatsu H, Schlis K, Davis KL, Martin PL, Nemecek ER, Yanik GA, Peters C, Baruchel A, Boissel N, Mechinaud F, Balduzzi A, Krueger J, June CH, Levine BL, Wood P, Taran T, Leung M, Mueller KT, Zhang Y, Sen K, Lebwohl D, Pulsipher MA, Grupp SA (2018) Tisagenlecleucel in children and Young adults with B-cell lymphoblastic leukemia. N Engl J Med 378:439–448

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  26. Schuster SJ, Bishop MR, Tam CS, Waller EK, Borchmann P, McGuirk JP, Jager U, Jaglowski S, Andreadis C, Westin JR, Fleury I, Bachanova V, Foley SR, Ho PJ, Mielke S, Magenau JM, Holte H, Pantano S, Pacaud LB, Awasthi R, Chu J, Anak O, Salles G, Maziarz RT, Investigators J (2019) Tisagenlecleucel in adult relapsed or refractory diffuse large B-cell lymphoma. N Engl J Med 380:45–56

    Article  CAS  PubMed  Google Scholar 

  27. Neelapu SS, Locke FL, Bartlett NL, Lekakis LJ, Miklos DB, Jacobson CA, Braunschweig I, Oluwole OO, Siddiqi T, Lin Y, Timmerman JM, Stiff PJ, Friedberg JW, Flinn IW, Goy A, Hill BT, Smith MR, Deol A, Farooq U, McSweeney P, Munoz J, Avivi I, Castro JE, Westin JR, Chavez JC, Ghobadi A, Komanduri KV, Levy R, Jacobsen ED, Witzig TE, Reagan P, Bot A, Rossi J, Navale L, Jiang Y, Aycock J, Elias M, Chang D, Wiezorek J, Go WY (2017) Axicabtagene ciloleucel CAR T-cell therapy in refractory large B-cell lymphoma. N Engl J Med 377:2531–2544

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  28. Bailey SR, Maus MV (2019) Gene editing for immune cell therapies. Nat Biotechnol [Epub ahead of print]

    Google Scholar 

  29. Xiao Q, Guo D, Chen S (2019) Application of CRISPR/Cas9-based gene editing in HIV-1/AIDS therapy. Front Cell Infect Microbiol 9:69

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  30. Zhang Y, Mu W, Wang H (2017) Gene editing in T cell therapy. J Genet Genomics 44:415–422

    Article  PubMed  Google Scholar 

  31. Perez EE, Wang J, Miller JC, Jouvenot Y, Kim KA, Liu O, Wang N, Lee G, Bartsevich VV, Lee YL, Guschin DY, Rupniewski I, Waite AJ, Carpenito C, Carroll RG, Orange JS, Urnov FD, Rebar EJ, Ando D, Gregory PD, Riley JL, Holmes MC, June CH (2008) Establishment of HIV-1 resistance in CD4+ T cells by genome editing using zinc-finger nucleases. Nat Biotechnol 26:808–816

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  32. Torikai H, Reik A, Liu PQ, Zhou Y, Zhang L, Maiti S, Huls H, Miller JC, Kebriaei P, Rabinovich B, Lee DA, Champlin RE, Bonini C, Naldini L, Rebar EJ, Gregory PD, Holmes MC, Cooper LJ (2012) A foundation for universal T-cell based immunotherapy: T cells engineered to express a CD19-specific chimeric-antigen-receptor and eliminate expression of endogenous TCR. Blood 119:5697–5705

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  33. Provasi E, Genovese P, Lombardo A, Magnani Z, Liu PQ, Reik A, Chu V, Paschon DE, Zhang L, Kuball J, Camisa B, Bondanza A, Casorati G, Ponzoni M, Ciceri F, Bordignon C, Greenberg PD, Holmes MC, Gregory PD, Naldini L, Bonini C (2012) Editing T cell specificity towards leukemia by zinc finger nucleases and lentiviral gene transfer. Nat Med 18:807–815

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  34. Torikai H, Reik A, Soldner F, Warren EH, Yuen C, Zhou Y, Crossland DL, Huls H, Littman N, Zhang Z, Tykodi SS, Kebriaei P, Lee DA, Miller JC, Rebar EJ, Holmes MC, Jaenisch R, Champlin RE, Gregory PD, Cooper LJ (2013) Toward eliminating HLA class I expression to generate universal cells from allogeneic donors. Blood 122:1341–1349

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  35. Berdien B, Mock U, Atanackovic D, Fehse B (2014) TALEN-mediated editing of endogenous T-cell receptors facilitates efficient reprogramming of T lymphocytes by lentiviral gene transfer. Gene Ther 21:539–548

    Article  CAS  PubMed  Google Scholar 

  36. Poirot L, Philip B, Schiffer-Mannioui C, Le Clerre D, Chion-Sotinel I, Derniame S, Potrel P, Bas C, Lemaire L, Galetto R, Lebuhotel C, Eyquem J, Cheung GW, Duclert A, Gouble A, Arnould S, Peggs K, Pule M, Scharenberg AM, Smith J (2015) Multiplex genome-edited T-cell manufacturing platform for "off-the-shelf" adoptive T-cell immunotherapies. Cancer Res 75:3853–3864

    Article  CAS  PubMed  Google Scholar 

  37. Valton J, Guyot V, Marechal A, Filhol JM, Juillerat A, Duclert A, Duchateau P, Poirot L (2015) A multidrug-resistant engineered CAR T cell for allogeneic combination immunotherapy. Mol Ther 23:1507–1518

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  38. Qasim W, Zhan H, Samarasinghe S, Adams S, Amrolia P, Stafford S, Butler K, Rivat C, Wright G, Somana K, Ghorashian S, Pinner D, Ahsan G, Gilmour K, Lucchini G, Inglott S, Mifsud W, Chiesa R, Peggs KS, Chan L, Farzeneh F, Thrasher AJ, Vora A, Pule M, Veys P (2017) Molecular remission of infant B-ALL after infusion of universal TALEN gene-edited CAR T cells. Sci Transl Med 9:eaaj2013

    Article  PubMed  Google Scholar 

  39. Schumann K, Lin S, Boyer E, Simeonov DR, Subramaniam M, Gate RE, Haliburton GE, Ye CJ, Bluestone JA, Doudna JA, Marson A (2015) Generation of knock-in primary human T cells using Cas9 ribonucleoproteins. Proc Natl Acad Sci U S A 112:10437–10442

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  40. Su S, Hu B, Shao J, Shen B, Du J, Du Y, Zhou J, Yu L, Zhang L, Chen F, Sha H, Cheng L, Meng F, Zou Z, Huang X, Liu B (2016) CRISPR-Cas9 mediated efficient PD-1 disruption on human primary T cells from cancer patients. Sci Rep 6:20070

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  41. Kim S, Kim D, Cho SW, Kim J, Kim J-S (2014) Highly efficient RNA-guided genome editing in human cells via delivery of purified Cas9 ribonucleoproteins. Genome Research 24(6):1012–1019

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  42. Ren J, Liu X, Fang C, Jiang S, June CH, Zhao Y (2017) Multiplex genome editing to generate universal CAR T cells resistant to PD1 inhibition. Clin Cancer Res 23:2255–2266

    Article  CAS  PubMed  Google Scholar 

  43. Liu X, Zhang Y, Cheng C, Cheng AW, Zhang X, Li N, Xia C, Wei X, Liu X, Wang H (2017) CRISPR-Cas9-mediated multiplex gene editing in CAR-T cells. Cell Res 27:154–157

    Article  PubMed  CAS  Google Scholar 

  44. Zhang W, Shi L, Zhao Z, Du P, Ye X, Li D, Cai Z, Han J, Cai J (2019) Disruption of CTLA-4 expression on peripheral blood CD8 + T cell enhances anti-tumor efficacy in bladder cancer. Cancer Chemother Pharmacol 83:911–920

    Article  CAS  PubMed  Google Scholar 

  45. Zhang Y, Zhang X, Cheng C, Mu W, Liu X, Li N, Wei X, Liu X, Xia C, Wang H (2017) CRISPR-Cas9 mediated LAG-3 disruption in CAR-T cells. Front Med 11:554–562

    Article  PubMed  Google Scholar 

  46. Rupp LJ, Schumann K, Roybal KT, Gate RE, Ye CJ, Lim WA, Marson A (2017) CRISPR/Cas9-mediated PD-1 disruption enhances anti-tumor efficacy of human chimeric antigen receptor T cells. Sci Rep 7:737

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  47. Jung IY, Kim YY, Yu HS, Lee M, Kim S, Lee J (2018) CRISPR/Cas9-mediated knockout of DGK improves antitumor activities of human T cells. Cancer Res 78:4692–4703

    Article  CAS  PubMed  Google Scholar 

  48. Cooper ML, Choi J, Staser K, Ritchey JK, Devenport JM, Eckardt K, Rettig MP, Wang B, Eissenberg LG, Ghobadi A, Gehrs LN, Prior JL, Achilefu S, Miller CA, Fronick CC, O’Neal J, Gao F, Weinstock DM, Gutierrez A, Fulton RS, DiPersio JF (2018) An “off-the-shelf” fratricide-resistant CAR-T for the treatment of T cell hematologic malignancies. Leukemia 32:1970–1983

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  49. Kim S, Koo T, Jee HG, Cho HY, Lee G, Lim DG, Shin HS, Kim JS (2018) CRISPR RNAs trigger innate immune responses in human cells. Genome Res 28:367–373.

    Article  CAS  PubMed Central  Google Scholar 

  50. Mu W, Tang N, Cheng C, Sun W, Wei X, Wang H (2019) In vitro transcribed sgRNA causes cell death by inducing interferon release. Protein Cell 10:461–465

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  51. Ren J, Zhang X, Liu X, Fang C, Jiang S, June CH, Zhao Y (2017) A versatile system for rapid multiplex genome-edited CAR T cell generation. Oncotarget 8:17002–17011

    PubMed  PubMed Central  Google Scholar 

  52. Lombardo A, Cesana D, Genovese P, Di Stefano B, Provasi E, Colombo DF, Neri M, Magnani Z, Cantore A, Lo Riso P, Damo M, Pello OM, Holmes MC, Gregory PD, Gritti A, Broccoli V, Bonini C, Naldini L (2011) Site-specific integration and tailoring of cassette design for sustainable gene transfer. Nat Methods 8:861–869

    Article  CAS  PubMed  Google Scholar 

  53. Sather BD, Ibarra GSR, Sommer K, Curinga G, Hale M, Khan IF, Singh S, Song YM, Gwiazda K, Sahni J, Jarjour J, Astrakhan A, Wagner TA, Scharenberg AM, Rawlings DJ (2015) Efficient modification of CCR5 in primary human hematopoietic cells using a megaTAL nuclease and AAV donor template. Sci Transl Med 7:307ra156

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  54. Eyquem J, Mansilla-Soto J, Giavridis T, van der Stegen SJ, Hamieh M, Cunanan KM, Odak A, Gonen M, Sadelain M (2017) Targeting a CAR to the TRAC locus with CRISPR/Cas9 enhances tumour rejection. Nature 543:113–117

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  55. MacLeod DT, Antony J, Martin AJ, Moser RJ, Hekele A, Wetzel KJ, Brown AE, Triggiano MA, Hux JA, Pham CD, Bartsevich VV, Turner CA, Lape J, Kirkland S, Beard CW, Smith J, Hirsch ML, Nicholson MG, Jantz D, McCreedy B (2017) Integration of a CD19 CAR into the TCR alpha chain locus streamlines production of allogeneic gene-edited CAR T cells. Mol Ther 25:949–961

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  56. Roth TL, Puig-Saus C, Yu R, Shifrut E, Carnevale J, Li PJ, Hiatt J, Saco J, Krystofinski P, Li H, Tobin V, Nguyen DN, Lee MR, Putnam AL, Ferris AL, Chen JW, Schickel JN, Pellerin L, Carmody D, Alkorta-Aranburu G, Del Gaudio D, Matsumoto H, Morell M, Mao Y, Cho M, Quadros RM, Gurumurthy CB, Smith B, Haugwitz M, Hughes SH, Weissman JS, Schumann K, Esensten JH, May AP, Ashworth A, Kupfer GM, Greeley SAW, Bacchetta R, Meffre E, Roncarolo MG, Romberg N, Herold KC, Ribas A, Leonetti MD, Marson A (2018) Reprogramming human T cell function and specificity with non-viral genome targeting. Nature 559:405–409

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  57. Dai X, Park JJ, Du Y, Kim HR, Wang G, Errami Y, Chen S (2019) One-step generation of modular CAR-T cells with AAV-Cpf1. Nat Methods 16:247–254

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  58. Berns K, Hijmans EM, Mullenders J, Brummelkamp TR, Velds A, Heimerikx M, Kerkhoven RM, Madiredjo M, Nijkamp W, Weigelt B, Agami R, Ge W, Cavet G, Linsley PS, Beijersbergen RL, Bernards R (2004) A large-scale RNAi screen in human cells identifies new components of the p53 pathway. Nature 428:431–437

    Article  CAS  PubMed  Google Scholar 

  59. Boutros M, Kiger AA, Armknecht S, Kerr K, Hild M, Koch B, Haas SA, Paro R, Perrimon N, Heidelberg Fly Array C (2004) Genome-wide RNAi analysis of growth and viability in drosophila cells. Science 303:832–835

    Article  CAS  PubMed  Google Scholar 

  60. Jackson AL, Burchard J, Schelter J, Chau BN, Cleary M, Lim L, Linsley PS (2006) Widespread siRNA "off-target" transcript silencing mediated by seed region sequence complementarity. RNA 12:1179–1187

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  61. Echeverri CJ, Beachy PA, Baum B, Boutros M, Buchholz F, Chanda SK, Downward J, Ellenberg J, Fraser AG, Hacohen N, Hahn WC, Jackson AL, Kiger A, Linsley PS, Lum L, Ma Y, Mathey-Prevot B, Root DE, Sabatini DM, Taipale J, Perrimon N, Bernards R (2006) Minimizing the risk of reporting false positives in large-scale RNAi screens. Nat Methods 3:777–779

    Article  CAS  PubMed  Google Scholar 

  62. Shalem O, Sanjana NE, Hartenian E, Shi X, Scott DA, Mikkelson T, Heckl D, Ebert BL, Root DE, Doench JG, Zhang F (2014) Genome-scale CRISPR-Cas9 knockout screening in human cells. Science 343:84–87

    Article  CAS  PubMed  Google Scholar 

  63. Wang T, Wei JJ, Sabatini DM, Lander ES (2014) Genetic screens in human cells using the CRISPR-Cas9 system. Science 343:80–84

    Article  CAS  PubMed  Google Scholar 

  64. Manguso RT, Pope HW, Zimmer MD, Brown FD, Yates KB, Miller BC, Collins NB, Bi K, LaFleur MW, Juneja VR, Weiss SA, Lo J, Fisher DE, Miao D, Van Allen E, Root DE, Sharpe AH, Doench JG, Haining WN (2017) In vivo CRISPR screening identifies Ptpn2 as a cancer immunotherapy target. Nature 547:413–418

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  65. Pan D, Kobayashi A, Jiang P, Ferrari de Andrade L, Tay RE, Luoma AM, Tsoucas D, Qiu X, Lim K, Rao P, Long HW, Yuan GC, Doench J, Brown M, Liu XS, Wucherpfennig KW (2018) A major chromatin regulator determines resistance of tumor cells to T cell-mediated killing. Science 359:770–775

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  66. Patel SJ, Sanjana NE, Kishton RJ, Eidizadeh A, Vodnala SK, Cam M, Gartner JJ, Jia L, Steinberg SM, Yamamoto TN, Merchant AS, Mehta GU, Chichura A, Shalem O, Tran E, Eil R, Sukumar M, Guijarro EP, Day CP, Robbins P, Feldman S, Merlino G, Zhang F, Restifo NP (2017) Identification of essential genes for cancer immunotherapy. Nature 548:537–542

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  67. Seki A, Rutz S (2018) Optimized RNP transfection for highly efficient CRISPR/Cas9-mediated gene knockout in primary T cells. J Exp Med 215:985–997

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  68. Shifrut E, Carnevale J, Tobin V, Roth TL, Woo JM, Bui CT, Li PJ, Diolaiti ME, Ashworth A, Marson A (2018) Genome-wide CRISPR screens in primary human T cells reveal key regulators of immune function. Cell 175(1958–1971):e15

    Google Scholar 

  69. Ting PY, Parker AE, Lee JS, Trussell C, Sharif O, Luna F, Federe G, Barnes SW, Walker JR, Vance J, Gao MY, Klock HE, Clarkson S, Russ C, Miraglia LJ, Cooke MP, Boitano AE, McNamara P, Lamb J, Schmedt C, Snead JL (2018) Guide swap enables genome-scale pooled CRISPR-Cas9 screening in human primary cells. Nat Methods 15:941–946

    Article  CAS  PubMed  Google Scholar 

  70. Veres A, Gosis BS, Ding Q, Collins R, Ragavendran A, Brand H, Erdin S, Cowan CA, Talkowski ME, Musunuru K (2014) Low incidence of off-target mutations in individual CRISPR-Cas9 and TALEN targeted human stem cell clones detected by whole-genome sequencing. Cell Stem Cell 15:27–30

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  71. Tan EP, Li Y, Velasco-Herrera Mdel C, Yusa K, Bradley A (2015) Off-target assessment of CRISPR-Cas9 guiding RNAs in human iPS and mouse ES cells. Genesis 53:225–236

    Article  CAS  PubMed  Google Scholar 

  72. Crosetto N, Mitra A, Silva MJ, Bienko M, Dojer N, Wang Q, Karaca E, Chiarle R, Skrzypczak M, Ginalski K, Pasero P, Rowicka M, Dikic I (2013) Nucleotide-resolution DNA double-strand break mapping by next-generation sequencing. Nat Methods 10:361–365

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  73. Tsai SQ, Zheng Z, Nguyen NT, Liebers M, Topkar VV, Thapar V, Wyvekens N, Khayter C, Iafrate AJ, Le LP, Aryee MJ, Joung JK (2015) GUIDE-seq enables genome-wide profiling of off-target cleavage by CRISPR-Cas nucleases. Nat Biotechnol 33:187–197

    Article  CAS  PubMed  Google Scholar 

  74. Kochenderfer JN, Feldman SA, Zhao Y, Xu H, Black MA, Morgan RA, Wilson WH, Rosenberg SA (2009) Construction and preclinical evaluation of an anti-CD19 chimeric antigen receptor. J Immunother 32:689–702

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  75. Carpenter RO, Evbuomwan MO, Pittaluga S, Rose JJ, Raffeld M, Yang S, Gress RE, Hakim FT, Kochenderfer JN (2013) B-cell maturation antigen is a promising target for adoptive T-cell therapy of multiple myeloma. Clin Cancer Res 19:2048–2060

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  76. Fraietta JA, Nobles CL, Sammons MA, Lundh S, Carty SA, Reich TJ, Cogdill AP, Morrissette JJD, DeNizio JE, Reddy S, Hwang Y, Gohil M, Kulikovskaya I, Nazimuddin F, Gupta M, Chen F, Everett JK, Alexander KA, Lin-Shiao E, Gee MH, Liu X, Young RM, Ambrose D, Wang Y, Xu J, Jordan MS, Marcucci KT, Levine BL, Garcia KC, Zhao Y, Kalos M, Porter DL, Kohli RM, Lacey SF, Berger SL, Bushman FD, June CH, Melenhorst JJ (2018) Disruption of TET2 promotes the therapeutic efficacy of CD19-targeted T cells. Nature 558:307–312

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  77. Saha S, Nakazawa Y, Huye LE, Doherty JE, Galvan DL, Rooney CM, Wilson MH (2012) piggyBac transposon system modification of primary human T cells. J Vis Exp 69:e4235

    Google Scholar 

  78. Monjezi R, Miskey C, Gogishvili T, Schleef M, Schmeer M, Einsele H, Ivics Z, Hudecek M (2017) Enhanced CAR T-cell engineering using non-viral sleeping beauty transposition from minicircle vectors. Leukemia 31:186–194

    Article  CAS  PubMed  Google Scholar 

  79. Kebriaei P, Izsvak Z, Narayanavari SA, Singh H, Ivics Z (2017) Gene therapy with the sleeping beauty transposon system. Trends Genet 33:852–870

    Article  CAS  PubMed  Google Scholar 

  80. Strecker J, Ladha A, Gardner Z, Schmid-Burgk JL, Makarova KS, Koonin EV, Zhang F (2019) RNA-guided DNA insertion with CRISPR-associated transposases. Science 365:48–53

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  81. Beane JD, Lee G, Zheng Z, Mendel M, Abate-Daga D, Bharathan M, Black M, Gandhi N, Yu Z, Chandran S, Giedlin M, Ando D, Miller J, Paschon D, Guschin D, Rebar EJ, Reik A, Holmes MC, Gregory PD, Restifo NP, Rosenberg SA, Morgan RA, Feldman SA (2015) Clinical scale zinc finger nuclease-mediated gene editing of PD-1 in tumor infiltrating lymphocytes for the treatment of metastatic melanoma. Mol Ther 23:1380–1390

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  82. Menger L, Sledzinska A, Bergerhoff K, Vargas FA, Smith J, Poirot L, Pule M, Hererro J, Peggs KS, Quezada SA (2016) TALEN-mediated inactivation of PD-1 in tumor-reactive lymphocytes promotes Intratumoral T-cell persistence and rejection of established Tumors. Cancer Res 76:2087–2093

    Article  CAS  PubMed  Google Scholar 

  83. Shi L, Meng T, Zhao Z, Han J, Zhang W, Gao F, Cai J (2017) CRISPR knock out CTLA-4 enhances the anti-tumor activity of cytotoxic T lymphocytes. Gene 636:36–41

    Article  CAS  PubMed  Google Scholar 

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Acknowledgements

H. W is supported by National Key Research and Development Program of China (2018YFA0107703), Strategic Priority Research Program of the Chinese Academy of Sciences (No. XDA16010503), and National Natural Science Foundation of China (No. 31722036). We apologize if we unintentionally missed citing some of the work that deserved citation.

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Author notes

  1. Xingying Zhang

    Present address: Institute for Stem Cell and Regeneration, Chinese Academy of Sciences, Beijing, China

  2. Chen Cheng

    Present address: School of Life Sciences, University of Science and Technology of China, Hefei, China

  3. Xingying Zhang and Chen Cheng contributed equally to this work.

Authors and Affiliations

  1. State Key Laboratory of Stem Cell and Reproductive Biology, Institute of Zoology, Chinese Academy of Sciences, Beijing, China

    Xingying Zhang, Chen Cheng, Wen Sun & Haoyi Wang

  2. University of Chinese Academy of Sciences, Beijing, China

    Xingying Zhang & Haoyi Wang

  3. Institute for Stem Cell and Regeneration, Chinese Academy of Sciences, Beijing, China

    Chen Cheng, Wen Sun & Haoyi Wang

Authors
  1. Xingying Zhang
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  2. Chen Cheng
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  3. Wen Sun
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  4. Haoyi Wang
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Corresponding author

Correspondence to Haoyi Wang .

Editor information

Editors and Affiliations

  1. Department of Cancer Immunology, Institute for Cancer Research, Oslo University Hospital-Radiumhospitalet, Oslo, Norway

    Mouldy Sioud

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Zhang, X., Cheng, C., Sun, W., Wang, H. (2020). Engineering T Cells Using CRISPR/Cas9 for Cancer Therapy. In: Sioud, M. (eds) RNA Interference and CRISPR Technologies. Methods in Molecular Biology, vol 2115. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-0290-4_23

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  • DOI: https://doi.org/10.1007/978-1-0716-0290-4_23

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  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-0716-0289-8

  • Online ISBN: 978-1-0716-0290-4

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