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Background: HIV pre-exposure prophylaxis (PrEP) has likely contributed to large decreases in HIV incidence among men who have sex with men (MSM) in several major cities. Berlin has seen a smaller decline, and affordable PrEP has been accessible through formal channels in Germany only since autumn 2017. We aimed to investigate knowledge and use of PrEP among MSM in Berlin, and factors predictive of a desire to use PrEP and history of PrEP use., Methods: Multicentre, paper-based, self-administered survey of adult MSM whose HIV status was negative or unknown at time of participation. Data were collected from 1 October 2017 to 2 April 2018., Results: 473 of 875 questionnaires were returned (response rate 54.1%; mean age 37.4 years, range 18-79). 90.0% of participants were aware of PrEP and, of these, 48.2% felt well informed about it. Among the 17.2% of participants reporting PrEP use, 59.3% indicated obtaining some or all of it from informal sources. 23.7% of those with no history of PrEP use reported having condomless anal intercourse (CAI) with two or more partners over the past six months. Worries about side effects, cost, not having a doctor who prescribes it, and a lack of information were the most frequently reported barriers to PrEP use. A desire to use PrEP and history of PrEP use were associated in our multivariable model with having multiple CAI partners. A history of PrEP use was associated with having a university degree, one or two parents born outside Germany, or friends living with HIV., Conclusions: We found high awareness of PrEP among MSM in Berlin, but also a strong need for more education on its pros, cons and proper use. The frequency of informal PrEP use was also high, raising urgent individual and public health concerns. Policy makers need to consider recent calls to improve access to PrEP and PrEP education through regular health services., Competing Interests: I have read the journal's policy and the authors of this manuscript have the following competing interests: The authors RNW, MG, JA, FK, RM, WO, DS, TW, CD, MS, and AN have declared that no competing interests exist. HJ received payments for research from Gilead Sciences GmbH; for Board membership from Gilead Sciences GmbH, MSD and ViiV Healthcare GmbH; for speaker activities from ViiV Healthcare GmbH, Abbvie Deutschland GmbH & Co. KG, Gilead Sciences GmbH, Gilead Sciences Ltd; and for travel/accommodation/meeting expenses from ViiV Healthcare GmbH, Abbvie Deutschland GmbH & Co. KG, Gilead Sciences GmbH, Gilead Sciences Ltd. SS declares that he received honoraria and support for lectures on the topic discussed in this paper by various commercial and non-commercial entities. The authors FK and KS are affiliated with Schwulenberatung Berlin gGmbH. The Schwulenberatung Berlin is not a commercial entity. Its gGmbH status allows it to accept private donations in addition to state funding. The funding of the Schwulenberatung Berlin comes primarily from the Berlin state government. Please see the following web link in English: http://www.schwulenberatungberlin.de/post.php?permalink=financing#seitenanfang. There are no patents, products in development or marketed products to declare. This does not alter our adherence to all the PLOS ONE policies on sharing data and materials.