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Rubius Therapeutics, Inc. – ‘10-K’ for 12/31/19 – ‘EX-101.DEF’

On:  Thursday, 3/12/20, at 9:01am ET   ·   For:  12/31/19   ·   Accession #:  1558370-20-2475   ·   File #:  1-38586

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  As Of               Filer                 Filing    For·On·As Docs:Size             Issuer                      Filing Agent

 3/12/20  Rubius Therapeutics, Inc.         10-K       12/31/19   86:13M                                    Toppan Merrill Bridge/FA

Annual Report   —   Form 10-K   —   Sect. 13 / 15(d) – SEA’34
Filing Table of Contents

Document/Exhibit                   Description                      Pages   Size 

 1: 10-K        Annual Report                                       HTML   1.72M 
 2: EX-4.1      Instrument Defining the Rights of Security Holders  HTML     59K 
 3: EX-10.9     Material Contract                                   HTML    120K 
 4: EX-21.1     Subsidiaries List                                   HTML     25K 
 5: EX-23.1     Consent of Experts or Counsel                       HTML     25K 
 6: EX-31.1     Certification -- §302 - SOA'02                      HTML     33K 
 7: EX-31.2     Certification -- §302 - SOA'02                      HTML     33K 
 8: EX-32.1     Certification -- §906 - SOA'02                      HTML     27K 
 9: EX-32.2     Certification -- §906 - SOA'02                      HTML     27K 
67: R1          Document and Entity Information                     HTML     62K 
22: R2          Consolidated Balance Sheets                         HTML    108K 
48: R3          Consolidated Balance Sheets (Parenthetical)         HTML     44K 
77: R4          Consolidated Statements of Operations and           HTML     74K 
                Comprehensive Loss                                               
65: R5          Consolidated Statements of Operations and           HTML     28K 
                Comprehensive Loss (Parenthetical)                               
21: R6          Consolidated Statements of Convertible Preferred    HTML    134K 
                Stock and Stockholders? Equity (Deficit)                         
47: R7          Consolidated Statements of Convertible Preferred    HTML     30K 
                Stock and Stockholders? Equity (Deficit)                         
                (Parenthetical)                                                  
75: R8          Consolidated Statements of Cash Flows               HTML    143K 
69: R9          Nature of the Business and Basis of Presentation    HTML     35K 
73: R10         Summary of Significant Accounting Policies          HTML    158K 
61: R11         Investments and Fair Value of Financial Assets and  HTML    173K 
                Liabilities                                                      
24: R12         Property, Plant and Equipment, Net                  HTML     55K 
51: R13         Accrued Expenses and Other Current Liabilities      HTML     46K 
74: R14         Debt                                                HTML     67K 
62: R15         Convertible Preferred Stock                         HTML     31K 
25: R16         Warrants to Purchase Convertible Preferred Stock    HTML     29K 
52: R17         Equity                                              HTML     32K 
72: R18         Stock-Based Compensation                            HTML    206K 
63: R19         Income Taxes                                        HTML    121K 
26: R20         Commitments and Contingencies                       HTML     34K 
35: R21         Leases                                              HTML    102K 
79: R22         Net Loss Per Share                                  HTML     85K 
53: R23         Related Parties                                     HTML     35K 
27: R24         Selected Quarterly Financial Data (Unaudited)       HTML    106K 
36: R25         Summary of Significant Accounting Policies          HTML    217K 
                (Policies)                                                       
80: R26         Summary of Significant Accounting Policies          HTML    125K 
                (Tables)                                                         
54: R27         Investments and Fair Value of Financial Assets and  HTML    174K 
                Liabilities (Tables)                                             
28: R28         Property, Plant and Equipment, Net (Tables)         HTML     51K 
34: R29         Accrued Expenses and Other Current Liabilities      HTML     46K 
                (Tables)                                                         
49: R30         Debt (Tables)                                       HTML     62K 
23: R31         Stock-Based Compensation (Tables)                   HTML    191K 
68: R32         Income Taxes (Tables)                               HTML    117K 
78: R33         Leases (Tables)                                     HTML     97K 
46: R34         Net Loss Per Share (Tables)                         HTML     86K 
20: R35         Selected Quarterly Financial Data (Unaudited)       HTML    106K 
                (Tables)                                                         
64: R36         Nature of the Business and Basis of Presentation    HTML     53K 
                (Details)                                                        
76: R37         Summary of Significant Accounting Policies          HTML     51K 
                (Details)                                                        
50: R38         Summary of Significant Accounting Policies -        HTML     36K 
                Estimated useful life (Details)                                  
19: R39         Summary of Significant Accounting Policies -        HTML    111K 
                Recently Adopted Accounting Pronouncements                       
                (Details)                                                        
40: R40         Investments and Fair Value of Financial Assets and  HTML     96K 
                Liabilities (Details)                                            
33: R41         Property, Plant and Equipment, Net (Details)        HTML     73K 
56: R42         Accrued Expenses and Other Current Liabilities      HTML     42K 
                (Details)                                                        
83: R43         Debt (Details)                                      HTML     87K 
39: R44         Debt - Current and Non current (Details)            HTML     39K 
32: R45         Convertible Preferred Stock (Details)               HTML     62K 
55: R46         Warrants to Purchase Convertible Preferred Stock    HTML     50K 
                (Details)                                                        
82: R47         Equity (Details)                                    HTML     70K 
41: R48         Stock-Based Compensation (Details)                  HTML     57K 
31: R49         Stock-Based Compensation - Employees, directors     HTML     34K 
                and non-employees (Details)                                      
16: R50         Stock-Based Compensation - Granted to               HTML     46K 
                non-employees, prior to the adoption of ASU                      
                2018-07 (Details)                                                
44: R51         Stock-Based Compensation - Option activity          HTML     80K 
                (Details)                                                        
70: R52         Stock-Based Compensation - Performance based stock  HTML     82K 
                options (Details)                                                
59: R53         Stock-Based Compensation - Restricted Common Stock  HTML    117K 
                (Details)                                                        
17: R54         Stock-Based Compensation - Compensation expense     HTML     72K 
                (Details)                                                        
45: R55         Income Taxes (Details)                              HTML     53K 
71: R56         Income Taxes - U S federal statutory income tax     HTML     49K 
                rate (Details)                                                   
60: R57         Income Taxes - Net deferred tax assets (Details)    HTML     60K 
18: R58         Income Taxes - Valuation allowance for deferred     HTML     29K 
                tax assets (Details)                                             
42: R59         Commitments and Contingencies - Collaborative       HTML     33K 
                Arrangements And Non-collaborative Arrangement                   
                (Details)                                                        
85: R60         Commitments and Contingencies - Defined             HTML     33K 
                Contribution Plan (Details)                                      
57: R61         Leases (Details)                                    HTML     61K 
29: R62         Leases - Minimum lease payments (Details)           HTML     67K 
37: R63         Leases - Sublease (Details)                         HTML     33K 
86: R64         Leases - Lease Portfolio (Details)                  HTML     50K 
58: R65         Net Loss Per Share - Weighted Average Shares        HTML     45K 
                (Details)                                                        
30: R66         Net Loss Per Share - Antidilutive Securities        HTML     42K 
                (Details)                                                        
38: R67         Related Parties (Details)                           HTML     84K 
84: R68         Selected Quarterly Financial Data (Unaudited)       HTML     37K 
                (Details)                                                        
43: XML         IDEA XML File -- Filing Summary                      XML    151K 
66: EXCEL       IDEA Workbook of Financial Reports                  XLSX    104K 
10: EX-101.INS  XBRL Instance -- ruby-20191231                       XML   2.67M 
12: EX-101.CAL  XBRL Calculations -- ruby-20191231_cal               XML    191K 
13: EX-101.DEF  XBRL Definitions -- ruby-20191231_def                XML    730K 
14: EX-101.LAB  XBRL Labels -- ruby-20191231_lab                     XML   1.64M 
15: EX-101.PRE  XBRL Presentations -- ruby-20191231_pre              XML   1.23M 
11: EX-101.SCH  XBRL Schema -- ruby-20191231                         XSD    197K 
81: ZIP         XBRL Zipped Folder -- 0001558370-20-002475-xbrl      Zip    213K 


‘EX-101.DEF’   —   XBRL Definitions — ruby-20191231_def


This Exhibit is an XBRL XML File.


                                                                                                                                                                                
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4 Subsequent Filings that Reference this Filing

  As Of               Filer                 Filing    For·On·As Docs:Size             Issuer                      Filing Agent

 2/27/23  Rubius Therapeutics, Inc.         10-K       12/31/22   77:7.2M
 6/08/22  Rubius Therapeutics, Inc.         S-3                    6:1M                                     Toppan Merrill/FA
 2/25/22  Rubius Therapeutics, Inc.         10-K       12/31/21   74:10M                                    Toppan Merrill Bridge/FA
 2/23/21  Rubius Therapeutics, Inc.         10-K       12/31/20   82:12M                                    Toppan Merrill Bridge/FA
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Filing Submission 0001558370-20-002475   –   Alternative Formats (Word / Rich Text, HTML, Plain Text, et al.)

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