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CORRESP
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CORRESP

 [ Janus Henderson Investors US LLC Letterhead ]

March 6, 2023

 VIA EDGAR

Samantha Brutlag

 Christina Fettig

Division of Investment Management

 U.S. Securities and Exchange
Commission

 100 F Street, N.E.

 Washington, DC 20549-0505

Re:
 JANUS INVESTMENT FUND (the “Registrant”)

1933 Act File No. 333-269480

Preliminary Proxy Statement/Prospectus on Form N-14

Dear Mses. Brutlag and Fettig:

 This letter responds to
comments provided by telephone on February 14, 2023 and February 27, 2023 with respect to the preliminary proxy statement/prospectus on Form N-14 (the “Registration Statement”) filed by the
Registrant related to the reorganization of Janus Henderson International Opportunities Fund (the “Target Fund”) with and into Janus Henderson Overseas Fund (the “Acquiring Fund,” and together, the “Funds”) (the
“Merger”), as filed with the Securities and Exchange Commission on January 31, 2023. The Staff’s comments, as we understand them, and the Registrant’s responses are below.

Comments Received on February 14, 2023

1.
 Staff Comment: Since the consent of the independent public accounting firm was not filed
with the Registration Statement, the Registration Statement cannot be automatically effective. Please amend the Registration Statement to include a delaying amendment in accordance with Rule 473 under the Securities Act of 1933, as amended.

 Response: A delaying amendment was filed on March 1, 2023. The Registrant notes that it
intends to formally request that the effectiveness of the Registration Statement be accelerated in order to maintain the planned timing of the Merger.

2.
 Staff Comment: Please provide hyperlinks to all documents that are incorporated by
reference into the Registration Statement.

 Response: The requested change has been made in all
relevant sections of the Registration Statement.

3.
 Staff Comment: Clarify what “substantially all” means in the below sentence, and
consider removing this disclosure:

 The Board of Trustees has approved a proposal to combine the Funds by having the
Target Fund transfer all or substantially all of its assets to the Acquiring Fund in exchange for shares of beneficial

interest of the Acquiring Fund and the assumption by the Acquiring Fund of all of the liabilities of the Target Fund.

Response: The reference to “substantially all” aligns with the definition of a “merger” under Rule 17a-8 under the Investment Company Act of 1940, as amended, upon which the Registrant is relying to effect the Merger. The reference to “substantially all” also is consistent with the requirements for a tax-free reorganization under Section 368 of the Internal Revenue Code of 1986, as amended. The Registrant believes that this disclosure is appropriate because it provides additional flexibility under
circumstances where certain assets cannot be transferred in connection with a merger. Accordingly, the Registrant has not removed this disclosure. For the Staff’s information, the Registrant notes that it currently anticipates that all assets
of the Target Fund will be transferred to the Acquiring Fund in connection with the Merger.

4.
 Staff Comment: In all instances where the disclosure discusses the potential benefits to
Janus Henderson Investors US LLC (the “Adviser”), please consider whether the Adviser having to waive less fees following the Merger could be a potential benefit to the Adviser.

Response: The Registrant has updated the disclosure as follows in all relevant sections of the Registration Statement
(additions underlined).

•

 potential benefits to the Adviser and its affiliates as a result of the Merger, and the Adviser’s belief
that while the Merger may result in some benefits and economies of scale for Janus Henderson and its affiliates, including, for example, an immaterial reduction in the level of operational expenses incurred for administrative and compliance services
as a result of the elimination of the Target Fund as a separate fund and, based on the fees and expenses of the Funds as of September 30, 2022, that the Adviser would be required to waive less of its advisory fees, the
Merger is not expected to result in a material change to the profitability of the Acquiring Fund or the Adviser.

5.
 Staff Comment: Please ensure that the list of expenses that are excluded from the Target
Fund’s and Acquiring Fund’s investment advisory fee waiver arrangements is aligned with the disclosure in the Funds’ prospectuses.

Response: The Registrant has updated the disclosure as follows in all relevant sections of the Registration Statement
(additions underlined).

 In addition, the Adviser has agreed to waive its investment advisory fee and/or reimburse certain other
Fund operating expenses to the extent that total annual operating expenses exceed 0.83% for the Target Fund, and 0.82% for the Acquiring Fund (excluding any performance adjustments to management fees, the fees payable pursuant to a
Rule 12b-1 plan, shareholder servicing fees, such as transfer agency fees (including out-of-pocket costs),
administrative services fees and any networking/omnibus fees payable by any share class, brokerage commissions, interest, dividends, taxes, acquired fund fees and expenses, and extraordinary expenses).

6.
 Staff Comment: Confirm that the disclosure regarding the pro forma net expense
ratios for certain share classes of the Acquiring Fund following the merger is accurate, as the pro forma net expense ratios for the Acquiring Fund in the Current and Pro Forma Fees and Expenses section of the Registration Statement
reflect that net expenses for all share classes will be the same or lower.

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 Response: The Registrant notes that the pro forma net expense ratios
for the Acquiring Fund in the Current and Pro Forma Fees and Expenses section of the Registration Statement reflect the application of the non-standard waiver arrangement wherein the Adviser has agreed
to waive the annual fund operating expenses of Class C Shares, Class R Shares, and Class D Shares to the extent they exceed 1.95%, 1.54%, and 0.95%, respectively.

7.
 Staff Comment: In the discussion of the U.S. federal income tax consequences of the Merger
on page 8 of the Registration Statement, please consider cross-referencing the subsequent Q&A regarding the estimated brokerage commissions or other transaction costs associated with the Merger for additional information about the repositioning.

 Response: The Registrant has updated the disclosure as follows (additions underlined).

If the Merger is approved, it is expected that approximately 75% of the portfolio holdings of the Target Fund will be sold by the Acquiring
Fund after the closing of the Merger. The Adviser anticipates that any sales of securities to align the portfolio holdings of the Target Fund with those of the Acquiring Fund following the Merger will result in a net capital loss to the Acquiring
Fund. Accordingly, the Adviser does not anticipate that the Merger will result in adverse tax consequences to shareholders of either of the Funds (as shareholders of the Acquiring Fund following the completion of the Merger). If, however, due to
market conditions or other factors, the Acquiring Fund experiences a net capital gain in connection with the disposition of securities following the Merger, shareholders of either of the Funds (as shareholders of the Acquiring Fund following the
completion of the Merger) will be taxed on any resulting capital gain distributions. Additional information about the repositioning of the Target Fund’s portfolio in connection with the Merger can be found under “Will either Fund pay
direct fees or expenses associated with the Merger?” below.

8.
 Staff Comment: In each instance, please disclose whether the Adviser will pay for the
direct fees and expenses associated with the Merger whether or not the Merger is consummated.

Response: The Registrant has updated the disclosure as follows in all relevant sections of the Registration Statement
(additions underlined).

 Janus Henderson will pay the direct fees and expenses associated with the Merger whether or not the
Merger is consummated, including preparation of the Proxy Statement/Prospectus, printing and mailing costs, and solicitation costs, which are estimated to be approximately between $334,000 and $506,000.

9.
 Staff Comment: In each instance, please disclose the percentage of the Target Fund’s
securities that will be sold in connection with the Merger.

 Response: The Registrant has updated
the disclosure as follows in all relevant sections of the Registration Statement (additions underlined).

 If the Merger is
approved, it is expected that approximately 75% of the portfolio holdings of the Target Fund will be sold by the Acquiring Fund after the closing of the Merger.

10.
 Staff Comment: Confirm that the fees presented in the fee tables represent current fees in
accordance with Item 3 of Form N-14.

 Response: The
Registrant so confirms.

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11.
 Staff Comment: Please revise the Current and Pro Forma Fees and Expenses section of
the Registration Statement to include footnotes regarding any expense limitation arrangements pursuant to the format prescribed in Item 3 of Form N-1A.

Response: The requested change has been made.

12.
 Staff Comment: In order to show the Fee Waiver line item in the fee table for the
Acquiring Fund, the expense limitation agreement must be in effect for at least one year following the effective date of the Registration Statement.

Response: The Registration Statement has been updated to reflect that the expense limitation agreement for the Acquiring
Fund will be in effect until March 31, 2024.

13.
 Staff Comment: Explain how the $557 figure in the below disclosure ties to the Statement
of Operations in the Acquiring Fund’s annual report for the period ended September 30, 2022, which reflects that $59,782 were reimbursed or waived.

For the fiscal year ended September 30, 2022, the Acquiring Fund’s investment advisory fee rate and fund operating expenses were
greater than the expense limit so $557 in fees or expenses were waived or reimbursed.

 Response: The Registrant
notes that the $557 figure represents the fees and expenses that were waived pursuant to the Acquiring Fund’s expense limitation agreement. The $59,782 figure in the Statement of Operations includes $59,225 of reimbursements to Class R
Shares, Class S Shares, and Class T Shares for administrative services fees. Such fees are excluded from the Acquiring Fund’s expense limitation agreement, but the Adviser reimburses the Acquired Fund
one-half of any administrative services fees that it retains for these Share Classes.

14.
 Staff Comment: Explain why the gross expense ratios for Class C Shares, Class S
Shares, and Class R Shares of the Target Fund presented in the Registration Statement do not align with the gross expense ratios for such Share Classes presented in the Financial Highlights for the Target Fund for the period ended
September 30, 2022.

 Response: The gross expense ratios for Class C Shares, Class S
Shares, and Class R Shares of the Target Fund presented in the Registration Statement reflect the maximum allowable 12b-1 fees for each Share Class. The gross expense ratios for Class C Shares,
Class S Shares, and Class R Shares presented in the Financial Highlights for the Target Fund for the period ended September 30, 2022 reflect 12b-1 fees that were accrued, a portion of which were
then waived.

15.
 Staff Comment: Please consider disclosing how long any fee waivers were factored into the
expense example calculations.

 Response: The Registrant has updated the disclosure as follows
(additions underlined, deletions stricken).

 The Examples also assume that your investment has a 5% return each
year and that the Funds’ operating expenses are equal to the Total Annual Fund Operating Expenses After Fee Waiver for the first year, as applicable, and the Total Annual Fund Operating Expenses thereafter remain the same as
shown above.

16.
 Staff Comment: Add a heading to page 15 of the Registration Statement to reflect that the
second part of the expense example reflects expenses if shares are not redeemed.

 Response: The
requested change has been made.

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17.
 Staff Comment: On page 30 of the Registration Statement, disclose the identity of the
accounting and performance survivor following the Merger.

 Response: The Registrant has updated the
disclosure as follows (additions underlined).

 The following information provides some indication of the risks of investing in each
Fund by showing how each Fund’s performance has varied over time. The Acquiring Fund, upon the completion of the Merger, will maintain the accounting history and performance track record of the Acquiring Fund.

18.
 Staff Comment: Disclose the dollar amount of capital loss carryforwards in the Target
Fund.

 Response: The disclosure has been updated to reflect that the Target Fund did not have any
capital loss carryforwards as of September 30, 2022.

   Comments Received on February 27, 2023

19.
 Staff Comment: If accurate, update the disclosure to reflect that the Merger is in the
best interest of the Funds and their shareholders [emphasis added] in all instances.

 Response:
The Registrant has reviewed the above-referenced language with counsel to the independent Trustees of the Registrant and together concluded that it does not need to be updated.

20.
 Staff Comment: In the Q&A on page 7 of the Registration Statement regarding whether
the Merger will result in higher Fund expenses, please clarify the answer as it relates to the Acquiring Fund’s annual operating expenses for certain share classes in recognition that the waiver arrangement will only be in place for a year
following the Merger.

 Response: The Registrant has updated the disclosure as follows (additions
underlined, deletions stricken).

 While the pro forma expense ratios for certain share
classes are anticipated to be higher than the expense ratios of the corresponding classes of the Funds, Tthe Adviser has agreed to cap the net annual fund operating expenses of Class C Shares, Class R Shares,
and Class D Shares of the Acquiring Fund following the Merger so they do not exceed the net annual fund operating expenses of each corresponding share class of each Fund as of September 30, 2022.

21.
 Staff Comment: Please include a side-by-side comparison of the fees and expenses of the Target Fund, the Acquiring Fund, and the pro forma expenses of the Acquiring Fund.

Response: The requested change has been made.

22.
 Staff Comment: Provide a comparison of the fundamental investment restrictions for the
Funds in the Registration Statement.

 Response: The requested change has been made and also is
provided in Appendix A attached hereto.

23.
 Staff Comment: Provide the Staff with a copy of the proxy card and the form of Opinion and
Consent of Counsel prior to the effective date of the Registration Statement.

 Response: The proxy
card and form of Opinion and Consent of Counsel are attached hereto as Appendix B and Appendix C, respectively.

24.
 Staff Comment: Include an undertaking in Item 17 of Part C to file the tax opinion upon
the completion of the Merger.

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 Response: The requested change has been made.

*        *        *

Please call me at (303) 394-7310 with any questions or comments.

Respectfully,

 /s/ Mary Clarke-Pearson

Mary Clarke-Pearson, Esq.

 Assistant Secretary to the Registrant

 Enclosure (via EDGAR)

cc:
 Abigail Murray, Esq.

Thea Kelley

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 Appendix A

Fundamental Investment Policies and Restrictions

 The
fundamental investment policies and restrictions for the Funds are identical, with the exception of the Funds’ fundamental policy on borrowing. The following is intended to compare the Funds’ fundamental investment policies and
restrictions. The Acquiring Fund will continue to have the same fundamental investment policies and rest